Material/Methoden, Ergebnisse, Diskussion und Einleitung vervollständigt
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@ -22,7 +22,7 @@ rm ./*.lot
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rm ./*.log
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rm ./*.out
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rm ./*.toc
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rm ./*.pdf
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# rm ./*.pdf
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#Entferne aus EPS-Dateien erstellte PDFs
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rm ./img/*-eps-converted-to.pdf
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style="font-size:19.77260017px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="495.3136 505.33832 509.80692 520.14789 529.10486"
|
||||
y="-312.15799"
|
||||
sodipodi:role="line"
|
||||
id="tspan44">2 Pyr</tspan><tspan
|
||||
style="font-size:19.77260017px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="375.4736 385.49832 389.96692 399.85333 409.4826 166.4666 176.4913 180.95992 192.40825 209.2545"
|
||||
y="-428.44601"
|
||||
sodipodi:role="line"
|
||||
id="tspan46">2 ATP2 AMP</tspan><tspan
|
||||
style="font-size:19.77260017px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="56.613598 66.638306 71.106903 81.329346 90.978378"
|
||||
y="-312.15799"
|
||||
sodipodi:role="line"
|
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id="tspan48">2 PEP</tspan></text>
|
||||
<text
|
||||
transform="scale(1,-1)"
|
||||
id="text50"
|
||||
x="106.5468"
|
||||
y="-413.466"><tspan
|
||||
style="font-size:19.77260017px;font-style:italic;font-variant:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O-Italic"
|
||||
x="106.5468 117.99513"
|
||||
y="-413.466"
|
||||
sodipodi:role="line"
|
||||
id="tspan52">AK</tspan></text>
|
||||
<text
|
||||
transform="scale(1,-1)"
|
||||
id="text54"
|
||||
x="29.507"
|
||||
y="-428.44611"><tspan
|
||||
style="font-size:19.77260017px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="29.507 39.531708 44.000317 55.44865 67.608803"
|
||||
y="-428.44611"
|
||||
sodipodi:role="line"
|
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id="tspan56">4 ADP</tspan><tspan
|
||||
style="font-size:19.77260017px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="123.6668 133.69151 138.16011 148.04654 157.6758"
|
||||
y="-462.68591"
|
||||
sodipodi:role="line"
|
||||
id="tspan58">2 ATP</tspan></text>
|
||||
<text
|
||||
transform="scale(1,-1)"
|
||||
id="text60"
|
||||
x="-1.1662"
|
||||
y="-124.5659"><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:bold;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O-Bold"
|
||||
x="-1.1662 7.8331599 16.95113 25.8319 33.661854 42.186649"
|
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y="-124.5659"
|
||||
sodipodi:role="line"
|
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id="tspan62">Konventionelle Glykolyse</tspan></text>
|
||||
<text
|
||||
transform="scale(1,-1)"
|
||||
id="text78"
|
||||
x="11.6875"
|
||||
y="-96.031403"><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="11.6875 16.873558 20.703783 29.296368 33.126595 41.617439 49.871067 58.633133"
|
||||
y="-96.031403"
|
||||
sodipodi:role="line"
|
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id="tspan80">- 2 ATP je Glukose</tspan><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="7.44102 15.881075 19.7113 28.303885 32.134109 40.607895 48.861523 57.623585 61.453812 65.504364 73.944412 77.774635 86.418205 95.180267 105.8744 115.68723"
|
||||
y="-75.962303"
|
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sodipodi:role="line"
|
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id="tspan82">+ 2 ATP je 3PGAK</tspan><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="7.44102 15.881075 19.7113 28.303885 32.134109 40.607895 48.861523 57.623585 61.453812 65.504364 73.944412 77.774635 86.58741 95.349472 105.77243"
|
||||
y="-55.511497"
|
||||
sodipodi:role="line"
|
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id="tspan84">+ 2 ATP je PPDK</tspan></text>
|
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<path
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|
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id="path86"
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style="fill:none;stroke:#000000;stroke-width:0.66310704;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:4;stroke-opacity:1;stroke-dasharray:none"
|
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d="m 0.262109,40.4789 191.214891,0" /><text
|
||||
transform="scale(1,-1)"
|
||||
id="text90"
|
||||
x="7.4503899"
|
||||
y="-19.985201"><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="7.4503899 15.890444 19.720671 28.313255 32.143482 40.617264 48.870892"
|
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y="-19.985201"
|
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sodipodi:role="line"
|
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id="tspan92">= 2 ATP</tspan></text>
|
||||
<text
|
||||
transform="scale(1,-1)"
|
||||
id="text94"
|
||||
x="244.42039"
|
||||
y="-125.5092"><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:bold;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O-Bold"
|
||||
x="244.42039 255.2162 259.38538 267.4187 275.11285 284.23083 288.40002 296.34854 303.11078 311.63556 315.46579 329.24442 333.4136 339.34537 343.1756 352.19189 361.25903 371.93622 381.20673 385.03696 396.98523 400.81543 409.88269 417.66177"
|
||||
y="-125.5092"
|
||||
sodipodi:role="line"
|
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id="tspan96">Glykolyse mit PPDK & PFK</tspan></text>
|
||||
<text
|
||||
transform="scale(1,-1)"
|
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id="text98"
|
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x="257.27469"
|
||||
y="-96.975998"><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="257.27469 262.46075 266.29099 274.88354 278.71378 287.20462 295.45825 304.22031 308.05054 312.1011 320.54114 324.37137 335.06549 338.99759 347.89523 355.04712 363.97864 370.60529"
|
||||
y="-96.975998"
|
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sodipodi:role="line"
|
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id="tspan100">- 1 ATP je Glukose</tspan><tspan
|
||||
style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
||||
x="253.02782 261.46786 265.2981 273.89069 277.72089 286.19467 294.4483 303.21036 307.04059 311.09116 319.53119 323.36142 332.00497 340.76706 351.46118 361.27402"
|
||||
y="-76.906502"
|
||||
sodipodi:role="line"
|
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id="tspan102">+ 2 ATP je 3PGAK</tspan><tspan
|
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style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
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x="253.02782 261.46786 265.2981 273.89069 277.72089 286.19467 294.4483 303.21036 307.04059 311.09116 319.53119 323.36142 333.17426"
|
||||
y="-56.4557"
|
||||
sodipodi:role="line"
|
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id="tspan104">+ 2 ATP je AK</tspan></text>
|
||||
<path
|
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id="path106"
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style="fill:none;stroke:#000000;stroke-width:0.66310704;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:4;stroke-opacity:1;stroke-dasharray:none"
|
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d="m 245.848,21.65 191.214,0" /><text
|
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transform="scale(1,-1)"
|
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id="text110"
|
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x="253.037"
|
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y="-1.15742"><tspan
|
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style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
|
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x="253.037 261.47705 265.30728 273.89987 277.7301 286.20386 294.45749"
|
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y="-1.15742"
|
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sodipodi:role="line"
|
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id="tspan112">= 5 ATP</tspan><tspan
|
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style="font-size:16.94790077px;font-variant:normal;font-weight:normal;writing-mode:lr-tb;fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Linux Biolinum O;-inkscape-font-specification:Linux Biolinum O"
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x="252.75027 261.19034 265.02057 273.61316 277.44336 285.91714 294.17078 302.93283 306.76306 310.8136 319.3045 323.13474 331.89679 340.65887 351.08182"
|
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y="-36.694122"
|
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sodipodi:role="line"
|
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id="tspan114">+ 2 ATP je PPDK</tspan></text>
|
||||
</g></svg>
|
||||
|
After Width: | Height: | Size: 15 KiB |
245
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Normal file
245
img/einleitung/ppdk_glycolysis.svg~
Normal file
|
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sodipodi:docname="ppdk_glycolysis.eps"><metadata
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d="m 837.23,4334.39 727.57,0"
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d="m 1108.75,4334.59 c 292.58,-0.2 362.11,185.9 362.11,185.9"
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d="m 1419.8,4481.5 64.3,74.81 -0.51,-98.63 c -13.63,20.89 -39.45,30.43 -63.79,23.82"
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id="g34"
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transform="scale(10,10)"><text
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transform="matrix(1,0,0,-1,273.467,295.052)"
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id="text36"><tspan
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style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-style:italic;font-variant:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri-Italic"
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x="0 10.2224342 20.4448684 32.6050174"
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id="tspan38">PPDK</tspan></text>
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<text
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transform="matrix(1,0,0,-1,182.8736,178.779)"
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id="text40"><tspan
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style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri"
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x="0 10.0247082 14.4933158 24.71575 34.9381842 196.836233 206.8609412 211.3295488 221.551983"
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y="0"
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sodipodi:role="line"
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id="tspan42">2 PPi2 Pi</tspan><tspan
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style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri"
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x="312.44 322.4647082 326.9333158 337.27425589 346.23124369"
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y="-133.379"
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sodipodi:role="line"
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id="tspan44">2 Pyr</tspan><tspan
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style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri"
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x="192.6 202.6247082 207.0933158 216.97973641 226.60899261 -16.407 -6.3822918 -1.9136842 9.5346512 26.3809064"
|
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y="-249.667"
|
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sodipodi:role="line"
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id="tspan46">2 ATP2 AMP</tspan><tspan
|
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style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri"
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x="-126.26 -116.2352918 -111.7666842 -101.54425 -91.8952212"
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y="-133.379"
|
||||
sodipodi:role="line"
|
||||
id="tspan48">2 PEP</tspan></text>
|
||||
<text
|
||||
transform="matrix(1,0,0,-1,106.5468,413.466)"
|
||||
id="text50"><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-style:italic;font-variant:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri-Italic"
|
||||
x="0 11.4483354"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan52">AK</tspan></text>
|
||||
<text
|
||||
transform="matrix(1,0,0,-1,29.507,428.4461)"
|
||||
id="text54"><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="0 10.0247082 14.4933158 25.9416512 38.1018002"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan56">4 ADP</tspan><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:19.7726;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="94.1598 104.1845082 108.6531158 118.53953641 128.16879261"
|
||||
y="-34.2398"
|
||||
sodipodi:role="line"
|
||||
id="tspan58">2 ATP</tspan></text>
|
||||
<text
|
||||
transform="matrix(1,0,0,-1,-1.1662,124.5659)"
|
||||
id="text60"><tspan
|
||||
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|
||||
x="0 8.99936032 18.11733052 26.99809961 34.82805314 43.35284684"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan62">Konven</tspan></text>
|
||||
</g><g
|
||||
id="g64"
|
||||
transform="matrix(88,0,0,-114,513,1358)"><rect
|
||||
x="0"
|
||||
y="0"
|
||||
width="1"
|
||||
height="1"
|
||||
transform="matrix(1,0,0,-1,0,1)"
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero"
|
||||
mask="url(#mask66)"
|
||||
id="rect70" /></g><g
|
||||
id="g72"
|
||||
transform="scale(10,10)"><text
|
||||
transform="matrix(1,0,0,-1,61.0613,124.565)"
|
||||
id="text74"><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:bold;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri-Bold"
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||||
x="0 9.1179702 18.2189925 26.7437862 30.9129696 35.082153 43.6069467 47.4880158 58.2838281 62.4530115 70.4863161 78.18047627 87.29844647 91.46762987 99.41616904 106.17838114"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan76">onelle Glykolyse</tspan></text>
|
||||
<text
|
||||
transform="matrix(1,0,0,-1,11.6875,96.0314)"
|
||||
id="text78"><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="0 5.1860574 9.0162828 17.6088681 21.4390935 29.92994056 38.18356786 46.94563216 50.77585756 59.67350506 68.11355926 74.02837636 77.85860176 85.84106266 89.72213176 98.61977926 105.67028429 114.60182759 121.22845649 129.66851069 133.49873609 141.93879029 150.66677236 156.34431886 162.36073489"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan80">- 2 ATP per glucose entry</tspan><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="-4.24648 4.1935742 8.0237996 16.6163849 20.4466103 28.92039421 37.17402151 45.93608581 49.76631121 53.81685931 62.25691351 66.08713891 74.73070671 83.49277101 94.18689591 103.99973001"
|
||||
y="20.0691"
|
||||
sodipodi:role="line"
|
||||
id="tspan82">+ 2 ATP je 3PGAK</tspan><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="-4.24648 4.1935742 8.0237996 16.6163849 20.4466103 28.92039421 37.17402151 45.93608581 49.76631121 53.81685931 62.25691351 66.08713891 74.89991116 83.66197546 94.08493396"
|
||||
y="40.5199"
|
||||
sodipodi:role="line"
|
||||
id="tspan84">+ 2 ATP je PPDK</tspan></text>
|
||||
</g><path
|
||||
d="m 2.62109,404.789 1912.14891,0"
|
||||
style="fill:none;stroke:#000000;stroke-opacity:1;stroke-width:6.63107;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:4;stroke-dasharray:none"
|
||||
id="path86" /><g
|
||||
id="g88"
|
||||
transform="scale(10,10)"><text
|
||||
transform="matrix(1,0,0,-1,7.45039,19.9852)"
|
||||
id="text90"><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="0 8.4400542 12.2702796 20.8628649 24.6930903 33.16687421 41.42050151"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan92">= 2 ATP</tspan></text>
|
||||
<text
|
||||
transform="matrix(1,0,0,-1,244.42039,125.5092)"
|
||||
id="text94"><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:bold;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri-Bold"
|
||||
x="0 10.7958123 14.9649957 22.9983003 30.69247234 39.81044254 43.97962594 51.92816511 58.69037721 67.21517091 71.04539631 84.82403901 88.99322241 94.92498741 98.75521281 107.77149561 116.83865719 127.51583419 136.78633549 140.61656089 152.56483039 156.39505579 165.46231499 173.24140109"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan96">Glykolyse mit PPDK & PFK</tspan></text>
|
||||
<text
|
||||
transform="matrix(1,0,0,-1,257.27469,96.976)"
|
||||
id="text98"><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="0 5.1860574 9.0162828 17.6088681 21.4390935 29.92993378 38.18356108 46.94562538 50.77585078 54.82639888 63.26645308 67.09667848 77.79080338 81.72289769 90.62054519 97.77241663 106.70395993 113.33058883"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan100">- 1 ATP je Glukose</tspan><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="-4.24688 4.1931742 8.0233996 16.6159849 20.4462103 28.91998574 37.17361304 45.93567734 49.76590274 53.81645084 62.25650504 66.08673044 74.73029214 83.49235644 94.18648134 103.99931544"
|
||||
y="20.0695"
|
||||
sodipodi:role="line"
|
||||
id="tspan102">+ 2 ATP je 3PGAK</tspan><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="-4.24688 4.1931742 8.0233996 16.6159849 20.4462103 28.91998574 37.17361304 45.93567734 49.76590274 53.81645084 62.25650504 66.08673044 75.89956454"
|
||||
y="40.5203"
|
||||
sodipodi:role="line"
|
||||
id="tspan104">+ 2 ATP je AK</tspan></text>
|
||||
</g><path
|
||||
d="m 2458.48,216.5 1912.14,0"
|
||||
style="fill:none;stroke:#000000;stroke-opacity:1;stroke-width:6.63107;stroke-linecap:butt;stroke-linejoin:miter;stroke-miterlimit:4;stroke-dasharray:none"
|
||||
id="path106" /><g
|
||||
id="g108"
|
||||
transform="scale(10,10)"><text
|
||||
transform="matrix(1,0,0,-1,253.037,1.15742)"
|
||||
id="text110"><tspan
|
||||
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|
||||
x="0 8.4400542 12.2702796 20.8628649 24.6930903 33.16686574 41.42049304"
|
||||
y="0"
|
||||
sodipodi:role="line"
|
||||
id="tspan112">= 5 ATP</tspan><tspan
|
||||
style="fill:#000000;fill-opacity:1;fill-rule:nonzero;stroke:none;font-family:Calibri;font-variant:normal;font-weight:normal;font-size:16.9479;writing-mode:lr;-inkscape-font-specification:Calibri"
|
||||
x="-0.286719 8.1533352 11.9835606 20.5761459 24.4063713 32.88014674 41.13377404 49.89583834 53.72606374 57.77661184 66.26750974 70.09773514 78.85979944 87.62186374 98.04482224"
|
||||
y="-35.5367"
|
||||
sodipodi:role="line"
|
||||
id="tspan114">+ 2 ATP je PPDK</tspan></text>
|
||||
</g></g></g></svg>
|
||||
|
After Width: | Height: | Size: 16 KiB |
BIN
img/einleitung/ppdk_hdr.png
Normal file
BIN
img/einleitung/ppdk_hdr.png
Normal file
Binary file not shown.
|
After Width: | Height: | Size: 1,006 KiB |
450
img/einleitung/swiveling_domain.eps
Normal file
450
img/einleitung/swiveling_domain.eps
Normal file
|
|
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|
|||
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||||
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|
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|
||||
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||||
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||||
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||||
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||||
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l 166.641 108.801 l 167.359 103.68 l 168 98.961 l 168.719 94.559 l 169.359
|
||||
90.078 l 170.078 86.078 l 170.719 82.238 l 171.441 78.398 l 172.078 75.121
|
||||
l 172.801 71.922 l 173.441 68.719 l 174.238 66.16 l 174.801 63.602 l 175.602
|
||||
61.039 l 176.16 58.879 l 176.961 56.719 l 177.52 54.879 l 178.32 53.762
|
||||
l 178.879 52.641 l 179.68 51.762 l 180.238 51.281 l 181.039 50.801 l 181.602
|
||||
50.559 l 182.961 50.559 l 183.762 50.719 l 184.32 50.961 l 185.121 51.199
|
||||
l 185.68 51.922 l 186.48 52.719 l 187.039 53.52 l 187.84 54.719 l 188.398
|
||||
56 l 189.199 57.281 l 189.762 58.719 l 190.559 60.16 l 191.121 61.68 l
|
||||
191.922 63.199 l 192.719 64.719 l 193.281 66.32 l 194.078 67.922 l 194.641
|
||||
69.52 l 195.441 70.961 l 196 72.238 l 196.801 73.602 l 197.359 75.121 l
|
||||
198.16 76.641 l 198.719 78.16 l 199.52 79.762 l 200.078 81.359 l 200.879
|
||||
83.039 l 201.441 84.641 l 202.238 86.32 l 202.801 87.84 l 203.602 89.199
|
||||
l 204.238 90.559 l 204.961 91.84 l 205.602 93.121 l 206.32 94.32 l 206.961
|
||||
95.602 l 207.68 96.879 l 208.32 98.16 l 209.039 99.602 l 209.68 100.961
|
||||
l 210.398 102.32 l 211.039 103.762 l 211.84 105.199 l 212.398 106.559 l
|
||||
213.199 108 l 213.762 109.359 l 214.559 110.719 l 215.121 111.84 l 215.922
|
||||
112.961 l 216.48 114 l 217.281 115.039 l 217.84 116.078 l 218.641 117.199
|
||||
l 219.199 118.48 l 220 119.68 l 220.559 120.879 l 221.359 122.16 l 221.922
|
||||
123.441 l 222.719 124.559 l 223.281 125.68 l 224.078 126.879 l 224.641
|
||||
127.84 l 225.441 128.801 l 226 129.762 l 226.801 130.801 l 227.359 131.84
|
||||
l 228.16 132.879 l 228.719 134 l 229.52 135.199 l 230.078 136.32 l 230.879
|
||||
137.52 l 231.441 138.641 l 232.238 139.84 l 233.039 141.039 l 233.602 142.238
|
||||
l 234.398 143.359 l 234.961 144.398 l 235.762 145.441 l 236.32 146.32 l
|
||||
237.121 147.199 l 237.68 148.16 l 238.48 149.199 l 239.039 150.238 l 239.84
|
||||
151.281 l 240.398 152.398 l 241.199 153.52 l 241.84 154.559 l 242.559 155.602
|
||||
l 243.199 156.641 l 243.922 157.602 l 244.559 158.48 l 245.281 159.359
|
||||
l 245.922 160.32 l 246.641 161.281 l 247.281 162.238 l 248 163.281 l 248.641
|
||||
164.32 l 249.441 165.359 l 250 166.48 l 250.801 167.602 l 251.359 168.641
|
||||
l 252.16 169.762 l 252.719 170.879 l 253.52 172 l 254.078 172.961 l 254.879
|
||||
174 l 255.441 174.879 l 256.238 175.762 l 256.801 176.641 l 257.602 177.52
|
||||
l 258.16 178.398 l 258.961 179.359 l 259.52 180.398 l 260.32 181.441 l
|
||||
260.879 182.48 l 261.68 183.441 l 262.238 184.48 l 263.039 185.441 l 263.602
|
||||
186.238 l 264.398 187.039 l 264.961 187.84 l 265.762 188.641 l 266.32 189.52
|
||||
l 267.121 190.32 l 267.68 191.199 l 268.48 192.16 l 269.039 193.039 l 269.84
|
||||
193.922 l 270.48 194.801 l 271.199 195.762 l 272 196.719 l 272.559 197.68
|
||||
l 273.359 198.48 l 273.922 199.281 l 274.719 200.078 l 275.281 200.801
|
||||
l 276.078 201.441 l 276.641 202.078 l 277.441 202.879 l 278 203.68 l 278.801
|
||||
204.48 l 279.441 205.359 l 280.16 206.238 l 280.801 207.121 l 281.52 207.922
|
||||
l 282.16 208.719 l 282.879 209.52 l 283.52 210.16 l 284.238 210.801 l 284.879
|
||||
211.52 l 285.602 212.238 l 286.238 213.039 l 287.039 213.602 l 287.602
|
||||
214.16 l 288.398 214.641 l 288.961 215.441 l 289.762 216.238 l 290.32 217.039
|
||||
l 291.121 217.84 l 291.68 218.719 l 292.48 219.602 l 293.039 220.398 l
|
||||
293.84 221.199 l 294.398 222 l 295.199 222.641 l 295.762 223.281 l 296.559
|
||||
224 l 297.121 224.801 l 297.922 225.52 l 298.48 226.32 l 299.281 227.121
|
||||
l 299.84 227.922 l 300.641 228.801 l 301.199 229.602 l 302 230.48 l 302.559
|
||||
231.121 l 303.359 231.84 l 303.922 232.559 l 304.719 233.281 l 305.281
|
||||
233.922 l 306.078 234.641 l 306.641 235.441 l 307.441 236.16 l 308.078 236.879
|
||||
l 308.801 237.68 l 309.441 238.48 l 310.16 239.281 l 310.801 240.078 l
|
||||
311.52 240.879 l 312.32 241.68 l 312.879 242.48 l 313.68 243.199 l 314.238
|
||||
243.84 l 315.039 244.48 l 315.602 245.121 l 316.398 245.84 l 317.039 246.48
|
||||
l 317.762 247.199 l 318.398 247.922 l 319.121 248.641 l 319.762 249.441
|
||||
l 320.48 250.16 l 321.121 250.961 l 321.84 251.602 l 322.48 252.238 l 323.199
|
||||
252.879 l 323.84 253.52 l 324.641 254.078 l 325.199 254.719 l 326 255.359
|
||||
l 326.559 256 l 327.359 256.641 l 327.922 257.359 l 328.719 258 l 329.281
|
||||
258.641 l 330.078 259.359 l 330.641 260.078 l 331.441 260.719 l 332 261.359
|
||||
l 332.801 262 l 333.359 262.559 l 334.16 263.121 l 334.719 263.602 l 335.52
|
||||
264.16 l 336.078 264.719 l 336.879 265.359 l 337.441 265.922 l 338.238
|
||||
266.559 l 338.801 267.121 l 339.602 267.762 l 340.16 268.32 l 340.961 268.961
|
||||
l 341.52 269.441 l 342.32 269.922 l 342.879 270.398 l 343.68 270.879 l
|
||||
344.238 271.441 l 345.039 271.922 l 345.68 272.48 l 346.398 272.961 l 347.039
|
||||
273.52 l 347.762 274 l 348.398 274.48 l 349.121 275.039 l 349.762 275.52
|
||||
l 350.48 276.078 l 351.121 276.559 l 351.84 277.039 l 352.641 277.52 l
|
||||
353.199 277.922 l 354 278.32 l 354.641 278.801 l 355.359 279.199 l 356 279.602
|
||||
l 356.719 280.078 l 357.359 280.48 l 358.078 280.961 l 358.719 281.359
|
||||
l 359.441 281.84 l 360.078 282.238 l 360.801 282.641 l 362.238 283.359 l
|
||||
362.801 283.68 l 363.602 284.078 l 364.16 284.48 l 364.961 284.801 l 365.52
|
||||
285.199 l 366.32 285.602 l 366.879 285.922 l 367.68 286.238 l 368.238 286.641
|
||||
l 369.039 286.961 l 369.602 287.359 l 370.398 287.762 l 370.961 288.078
|
||||
l 371.762 288.398 l 372.32 288.719 l 373.121 289.039 l 373.68 289.281 l
|
||||
374.48 289.602 l 375.039 289.84 l 375.84 290.16 l 376.398 290.48 l 377.199
|
||||
290.879 l 377.762 291.199 l 378.559 291.441 l 379.121 291.762 l 379.922
|
||||
292.078 l 380.48 292.32 l 381.281 292.559 l 381.84 292.719 l 382.641 293.039
|
||||
l 383.281 293.281 l 384 293.52 l 384.641 293.762 l 385.359 294.078 l 386
|
||||
294.32 l 386.719 294.559 l 387.359 294.801 l 388.078 295.039 l 388.719
|
||||
295.359 l 389.441 295.602 l 390.078 295.84 l 390.801 296.078 l 391.441 296.238
|
||||
l 392.238 296.48 l 392.961 296.641 l 393.602 296.879 l 394.32 297.039 l
|
||||
394.961 297.281 l 395.68 297.52 l 396.32 297.68 l 397.039 297.922 l 397.68
|
||||
298.16 l 398.398 298.32 l 399.039 298.559 l 399.84 298.719 l 400.398 298.879
|
||||
l 401.199 299.039 l 401.762 299.199 l 402.559 299.441 l 403.121 299.602
|
||||
l 403.922 299.762 l 404.48 299.922 l 405.281 300.078 l 405.84 300.32 l
|
||||
406.641 300.48 l 407.199 300.641 l 408 300.801 l 408.559 301.039 l 409.359
|
||||
301.199 l 409.922 301.359 l 410.719 301.52 l 411.281 301.602 l 412.078
|
||||
301.762 l 412.641 301.922 l 413.441 302 l 414 302.16 l 414.801 302.32 l
|
||||
415.359 302.48 l 416.16 302.641 l 416.719 302.801 l 417.52 302.879 l 418.078
|
||||
303.039 l 418.879 303.199 l 419.441 303.281 l 420.238 303.359 l 420.879
|
||||
303.52 l 421.602 303.602 l 422.238 303.68 l 422.961 303.84 l 423.602 303.922
|
||||
l 424.32 304.078 l 424.961 304.16 l 425.68 304.32 l 426.32 304.398 l 427.039
|
||||
304.559 l 427.68 304.641 l 428.398 304.801 l 429.039 304.879 l 429.84 304.961
|
||||
l 430.398 305.039 l 431.199 305.199 l 431.762 305.281 l 432.559 305.359
|
||||
l 433.281 305.441 l 433.922 305.52 l 434.641 305.68 l 435.281 305.762 l
|
||||
436 305.922 l 436.641 306 l 437.441 306.078 l 438 306.16 l 438.801 306.238
|
||||
l 439.359 306.32 l 440.16 306.32 l 440.719 306.398 l 441.52 306.48 l 442.078
|
||||
306.641 l 442.879 306.719 l 443.441 306.801 l 444.238 306.879 l 444.801
|
||||
306.961 l 445.602 307.039 l 446.16 307.121 l 446.961 307.199 l 447.52 307.281
|
||||
l 448.32 307.359 l 448.879 307.441 l 449.68 307.52 l 450.238 307.52 l 451.039
|
||||
307.602 l 451.602 307.68 l 452.398 307.762 l 452.961 307.84 l 453.762 307.922
|
||||
l 454.32 308 l 455.121 308.078 l 455.68 308.16 l 456.48 308.16 l 457.039
|
||||
308.238 l 457.84 308.238 l 458.48 308.32 l 459.199 308.398 l 459.84 308.48
|
||||
l 460.008 308.48 l S Q
|
||||
0 g
|
||||
BT
|
||||
12 0 0 12 305.679375 338.24 Tm
|
||||
/f-0-0 1 Tf
|
||||
[(Fraktion 2 \(200 mM\))]TJ
|
||||
ET
|
||||
0 0 1 rg
|
||||
q 1 0 0 -1 0 384 cm
|
||||
412.961 42.16 m 446.719 42.16 l 50.879 315.281 m 70.719 315.281 l 71.359
|
||||
315.199 l 75.441 315.199 l 76.238 315.121 l 76.801 315.121 l 77.602 315.039
|
||||
l 78.16 315.039 l 78.961 314.961 l 79.52 314.879 l 80.32 314.879 l 80.879
|
||||
314.801 l 81.68 314.641 l 82.238 314.559 l 83.039 314.48 l 83.602 314.32
|
||||
l 84.398 314.078 l 84.961 313.922 l 85.762 313.762 l 86.32 313.52 l 87.121
|
||||
313.281 l 87.68 313.121 l 88.48 312.879 l 89.039 312.641 l 89.84 312.398
|
||||
l 90.398 312.238 l 91.199 312 l 91.762 311.84 l 92.559 311.68 l 93.121
|
||||
311.52 l 93.922 311.359 l 94.48 311.281 l 95.281 311.199 l 95.922 311.121
|
||||
l 96.641 311.039 l 97.281 311.039 l 98 310.961 l 99.359 310.961 l 100.16
|
||||
310.879 l 102.078 310.879 l 102.879 310.801 l 104.238 310.801 l 104.879
|
||||
310.719 l 105.602 310.719 l 106.238 310.641 l 106.961 310.641 l 107.602
|
||||
310.559 l 108.32 310.559 l 108.961 310.48 l 109.68 310.48 l 110.32 310.398
|
||||
l 111.039 310.32 l 111.68 310.32 l 112.48 310.238 l 113.039 310.16 l 113.84
|
||||
310.16 l 114.398 310.078 l 115.199 310 l 115.762 310 l 116.559 309.922
|
||||
l 117.121 309.84 l 117.922 309.84 l 118.48 309.762 l 119.281 309.68 l 119.84
|
||||
309.68 l 120.641 309.602 l 121.199 309.602 l 122 309.52 l 122.559 309.52
|
||||
l 123.359 309.441 l 123.922 309.441 l 124.719 309.359 l 125.281 309.281
|
||||
l 126.078 309.199 l 126.641 309.121 l 127.441 309.121 l 128 309.039 l 128.801
|
||||
308.961 l 129.359 308.879 l 130.16 308.879 l 130.719 308.801 l 131.52 308.719
|
||||
l 132.078 308.559 l 132.879 308.48 l 133.52 308.32 l 134.238 308.16 l 134.879
|
||||
307.922 l 135.602 307.762 l 136.238 307.441 l 136.961 307.199 l 137.602
|
||||
306.879 l 138.32 306.48 l 139.121 306.078 l 139.68 305.68 l 140.48 305.121
|
||||
l 141.121 304.48 l 141.84 303.922 l 142.48 303.121 l 143.199 302.238 l
|
||||
143.84 301.359 l 144.559 300.32 l 145.199 299.281 l 145.922 298.078 l 146.559
|
||||
296.719 l 147.281 295.441 l 147.922 293.922 l 148.641 292.398 l 149.281
|
||||
290.801 l 150.078 289.039 l 150.641 287.199 l 151.441 285.359 l 152 283.121
|
||||
l 152.801 280.879 l 153.359 278.559 l 154.16 276.16 l 154.719 273.68 l
|
||||
155.52 271.121 l 156.078 268.48 l 156.879 265.84 l 157.441 263.121 l 158.238
|
||||
260.32 l 158.801 257.52 l 159.602 254.641 l 160.16 251.602 l 160.961 248.559
|
||||
l 161.52 245.52 l 162.32 242.559 l 162.879 239.52 l 163.68 236.641 l 164.238
|
||||
233.762 l 165.039 230.961 l 165.602 228.238 l 166.398 225.602 l 166.961
|
||||
222.961 l 167.762 220.719 l 168.32 218.398 l 169.121 216.16 l 169.68 214.16
|
||||
l 170.48 212.16 l 171.121 210.16 l 171.84 208.16 l 172.48 206.238 l 173.199
|
||||
204.641 l 173.84 203.441 l 174.559 202.238 l 175.199 201.359 l 175.922
|
||||
200.641 l 176.559 199.922 l 177.281 199.52 l 178.719 198.801 l 179.441 198.398
|
||||
l 180.078 197.922 l 180.801 197.602 l 181.441 197.84 l 182.16 198 l 182.801
|
||||
198.398 l 183.52 199.199 l 184.16 200 l 184.879 200.961 l 185.52 202.078
|
||||
l 186.238 203.199 l 186.879 204.48 l 187.68 205.762 l 188.238 207.039 l
|
||||
189.039 208.16 l 189.602 209.121 l 190.398 210.078 l 190.961 210.641 l
|
||||
191.762 211.121 l 192.32 211.602 l 193.121 212.961 l 193.68 214.238 l 194.48
|
||||
215.602 l 195.039 216.961 l 195.84 218.238 l 196.398 219.52 l 197.199 220.719
|
||||
l 197.762 221.84 l 198.559 222.48 l 199.121 222.641 l 199.922 222.719 l
|
||||
200.48 223.199 l 201.281 223.922 l 201.84 224.719 l 202.641 225.52 l 203.199
|
||||
226.398 l 204 227.199 l 204.559 228 l 205.359 228.801 l 205.922 229.602
|
||||
l 206.719 230.32 l 207.281 231.121 l 208.078 231.762 l 208.719 232 l 209.441
|
||||
232.238 l 210.078 232.398 l 210.801 232.238 l 211.441 232.16 l 212.16 232.32
|
||||
l 212.801 232.879 l 213.52 233.359 l 214.16 233.762 l 214.879 234.16 l
|
||||
215.52 234.48 l 216.32 234.719 l 216.879 234.961 l 217.68 235.199 l 218.398
|
||||
234.879 l 219.039 234.559 l 219.762 234.238 l 220.398 234.238 l 221.121
|
||||
234.32 l 221.762 234.32 l 222.48 234.48 l 223.121 234.641 l 223.84 234.719
|
||||
l 224.48 234.879 l 225.281 234.961 l 225.84 235.199 l 226.641 235.441 l
|
||||
227.199 235.68 l 228 235.602 l 228.559 235.359 l 229.359 235.039 l 229.922
|
||||
234.801 l 230.719 234.559 l 231.281 234.32 l 232.641 234.32 l 233.441 234.238
|
||||
l 234 234.16 l 234.801 234.078 l 235.359 234 l 236.16 233.84 l 236.719
|
||||
233.602 l 237.52 233.359 l 238.078 232.879 l 238.879 232.398 l 239.441 232
|
||||
l 240.238 231.68 l 240.801 231.441 l 241.602 231.281 l 242.16 231.199 l
|
||||
242.961 231.121 l 244.32 231.121 l 244.879 231.199 l 245.68 231.281 l 246.32
|
||||
231.441 l 247.039 231.52 l 247.68 231.199 l 248.398 230.961 l 249.039 230.641
|
||||
l 249.762 230.238 l 250.398 229.922 l 251.121 229.68 l 251.762 229.602
|
||||
l 252.48 229.441 l 253.121 229.441 l 253.922 229.359 l 255.281 229.359 l
|
||||
255.84 229.281 l 256.641 229.281 l 257.199 228.801 l 258 228.238 l 258.719
|
||||
227.68 l 259.359 227.359 l 260.078 227.121 l 260.719 226.801 l 261.441
|
||||
226.719 l 262.078 226.559 l 262.879 226.559 l 263.441 226.641 l 264.238
|
||||
226.719 l 264.801 226.961 l 265.602 227.199 l 266.16 227.52 l 266.961 227.52
|
||||
l 267.52 227.441 l 268.32 227.359 l 270.238 227.359 l 271.039 227.441 l
|
||||
271.602 227.602 l 272.398 227.762 l 272.961 228.238 l 273.762 228.641 l
|
||||
274.32 229.121 l 275.121 229.602 l 275.68 230.078 l 276.48 230.398 l 277.039
|
||||
230.641 l 277.84 230.879 l 278.398 231.199 l 279.199 231.68 l 279.762 232.078
|
||||
l 280.559 232.641 l 281.121 233.281 l 281.922 233.922 l 282.48 234.719
|
||||
l 283.281 235.441 l 283.922 236.238 l 284.641 237.121 l 285.281 237.922
|
||||
l 286 238.719 l 286.641 239.441 l 287.359 240.16 l 288 240.879 l 288.719
|
||||
241.52 l 289.359 242.16 l 290.078 242.801 l 290.719 243.441 l 291.52 244.078
|
||||
l 292.078 244.879 l 292.879 245.68 l 293.441 246.48 l 294.238 247.199 l
|
||||
294.801 248 l 295.602 248.801 l 296.16 249.441 l 296.961 250.16 l 297.52
|
||||
250.801 l 298.32 251.359 l 299.039 252 l 299.68 252.641 l 300.48 253.359
|
||||
l 301.039 254.078 l 301.84 254.879 l 302.398 255.68 l 303.199 256.398 l
|
||||
303.762 257.199 l 304.559 258 l 305.121 258.801 l 305.922 259.602 l 306.48
|
||||
260.32 l 307.281 261.039 l 307.84 261.68 l 308.641 262.32 l 309.199 262.961
|
||||
l 310 263.52 l 310.559 264.16 l 311.359 264.719 l 311.922 265.441 l 312.719
|
||||
266.16 l 313.281 266.879 l 314.078 267.602 l 314.641 268.32 l 315.441 268.961
|
||||
l 316 269.602 l 316.801 270.16 l 317.359 270.801 l 318.16 271.281 l 318.719
|
||||
271.84 l 319.52 272.398 l 320.078 272.961 l 320.879 273.602 l 321.52 274.16
|
||||
l 322.238 274.801 l 322.879 275.359 l 323.602 276 l 324.238 276.559 l 324.961
|
||||
277.199 l 325.602 277.762 l 326.32 278.398 l 326.961 278.961 l 327.68 279.441
|
||||
l 328.32 279.922 l 329.121 280.398 l 329.68 280.879 l 330.48 281.281 l
|
||||
331.039 281.84 l 331.84 282.32 l 332.398 282.879 l 333.199 283.359 l 333.762
|
||||
283.922 l 334.559 284.398 l 335.121 284.879 l 335.922 285.359 l 336.48
|
||||
285.84 l 337.281 286.238 l 337.84 286.641 l 338.641 287.039 l 339.441 287.52
|
||||
l 340 287.922 l 340.801 288.398 l 341.359 288.801 l 342.16 289.199 l 342.719
|
||||
289.602 l 343.52 290 l 344.078 290.48 l 344.879 290.879 l 345.441 291.281
|
||||
l 346.238 291.762 l 346.801 292.078 l 347.602 292.48 l 348.16 292.801 l
|
||||
348.961 293.121 l 349.52 293.441 l 350.32 293.762 l 350.879 294.16 l 351.68
|
||||
294.48 l 352.238 294.801 l 353.039 295.199 l 353.602 295.52 l 354.398 295.922
|
||||
l 354.961 296.238 l 355.762 296.559 l 356.32 296.879 l 357.121 297.121
|
||||
l 357.762 297.359 l 358.48 297.68 l 359.121 297.922 l 359.84 298.238 l 360.48
|
||||
298.48 l 361.199 298.801 l 361.84 299.121 l 362.559 299.359 l 363.199 299.602
|
||||
l 363.922 299.922 l 364.559 300.238 l 365.281 300.48 l 365.922 300.801
|
||||
l 366.719 301.039 l 367.281 301.281 l 368.078 301.441 l 368.641 301.602
|
||||
l 369.441 301.68 l 370 301.922 l 370.801 302.16 l 371.359 302.32 l 372.16
|
||||
302.559 l 372.719 302.801 l 373.52 303.039 l 374.078 303.281 l 374.879
|
||||
303.52 l 375.441 303.762 l 376.238 303.922 l 376.801 304.16 l 377.602 304.32
|
||||
l 378.398 304.48 l 378.961 304.641 l 379.762 304.879 l 380.32 305.039 l
|
||||
381.121 305.199 l 381.68 305.359 l 382.48 305.602 l 383.039 305.762 l 383.84
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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||||
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|
||||
/f-0-0 1 Tf
|
||||
[(-50)]TJ
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
57.52 305.441 m 61.121 305.441 l 57.52 273.039 m 64.727 273.039 l S Q
|
||||
BT
|
||||
12 0 0 12 45.301875 107.36 Tm
|
||||
/f-0-0 1 Tf
|
||||
(0)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
57.52 240.559 m 61.121 240.559 l 57.52 208.078 m 64.727 208.078 l S Q
|
||||
BT
|
||||
12 0 0 12 39.72375 172.32 Tm
|
||||
/f-0-0 1 Tf
|
||||
(50)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
57.52 175.602 m 61.121 175.602 l 57.52 143.199 m 64.727 143.199 l S Q
|
||||
BT
|
||||
12 0 0 12 34.145625 237.2 Tm
|
||||
/f-0-0 1 Tf
|
||||
(100)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
57.52 110.719 m 61.121 110.719 l 57.52 78.238 m 64.727 78.238 l S Q
|
||||
BT
|
||||
12 0 0 12 34.145625 302.16 Tm
|
||||
/f-0-0 1 Tf
|
||||
(150)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
57.52 45.84 m 61.121 45.84 l 57.52 13.359 m 64.727 13.359 l S Q
|
||||
BT
|
||||
12 0 0 12 34.145625 367.04 Tm
|
||||
/f-0-0 1 Tf
|
||||
(200)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
57.52 337.922 m 57.52 330.719 l 57.52 13.359 m 57.52 20.559 l 57.527 20.559
|
||||
l S Q
|
||||
BT
|
||||
12 0 0 12 54.730938 28.08 Tm
|
||||
/f-0-0 1 Tf
|
||||
(0)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
66.719 337.922 m 66.719 334.32 l 66.719 13.359 m 66.719 16.961 l 75.84
|
||||
337.922 m 75.84 334.32 l 75.84 13.359 m 75.84 16.961 l 85.039 337.922 m
|
||||
85.039 334.32 l 85.039 13.359 m 85.039 16.961 l 94.16 337.922 m 94.16 334.32
|
||||
l 94.16 13.359 m 94.16 16.961 l 103.359 337.922 m 103.359 334.32 l 103.359
|
||||
13.359 m 103.359 16.961 l 112.48 337.922 m 112.48 334.32 l 112.48 13.359
|
||||
m 112.48 16.961 l 121.68 337.922 m 121.68 334.32 l 121.68 13.359 m 121.68
|
||||
16.961 l 130.801 337.922 m 130.801 334.32 l 130.801 13.359 m 130.801 16.961
|
||||
l 140 337.922 m 140 334.32 l 140 13.359 m 140 16.961 l 149.199 337.922
|
||||
m 149.199 330.719 l 149.199 13.359 m 149.199 20.559 l 149.207 20.559 l S Q
|
||||
BT
|
||||
12 0 0 12 140.832813 28.08 Tm
|
||||
/f-0-0 1 Tf
|
||||
(100)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
158.32 337.922 m 158.32 334.32 l 158.32 13.359 m 158.32 16.961 l 167.52
|
||||
337.922 m 167.52 334.32 l 167.52 13.359 m 167.52 16.961 l 176.641 337.922
|
||||
m 176.641 334.32 l 176.641 13.359 m 176.641 16.961 l 185.84 337.922 m 185.84
|
||||
334.32 l 185.84 13.359 m 185.84 16.961 l 194.961 337.922 m 194.961 334.32
|
||||
l 194.961 13.359 m 194.961 16.961 l 204.16 337.922 m 204.16 334.32 l 204.16
|
||||
13.359 m 204.16 16.961 l 213.281 337.922 m 213.281 334.32 l 213.281 13.359
|
||||
m 213.281 16.961 l 222.48 337.922 m 222.48 334.32 l 222.48 13.359 m 222.48
|
||||
16.961 l 231.68 337.922 m 231.68 334.32 l 231.68 13.359 m 231.68 16.961
|
||||
l 240.801 337.922 m 240.801 330.719 l 240.801 13.359 m 240.801 20.559 l
|
||||
240.809 20.559 l S Q
|
||||
BT
|
||||
12 0 0 12 232.432813 28.08 Tm
|
||||
/f-0-0 1 Tf
|
||||
(200)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
250 337.922 m 250 334.32 l 250 13.359 m 250 16.961 l 259.121 337.922 m
|
||||
259.121 334.32 l 259.121 13.359 m 259.121 16.961 l 268.32 337.922 m 268.32
|
||||
334.32 l 268.32 13.359 m 268.32 16.961 l 277.441 337.922 m 277.441 334.32
|
||||
l 277.441 13.359 m 277.441 16.961 l 286.641 337.922 m 286.641 334.32 l
|
||||
286.641 13.359 m 286.641 16.961 l 295.762 337.922 m 295.762 334.32 l 295.762
|
||||
13.359 m 295.762 16.961 l 304.961 337.922 m 304.961 334.32 l 304.961 13.359
|
||||
m 304.961 16.961 l 314.078 337.922 m 314.078 334.32 l 314.078 13.359 m
|
||||
314.078 16.961 l 323.281 337.922 m 323.281 334.32 l 323.281 13.359 m 323.281
|
||||
16.961 l 332.48 337.922 m 332.48 330.719 l 332.48 13.359 m 332.48 20.559
|
||||
l 332.488 20.559 l S Q
|
||||
BT
|
||||
12 0 0 12 324.112813 28.08 Tm
|
||||
/f-0-0 1 Tf
|
||||
(300)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
341.602 337.922 m 341.602 334.32 l 341.602 13.359 m 341.602 16.961 l 350.801
|
||||
337.922 m 350.801 334.32 l 350.801 13.359 m 350.801 16.961 l 359.922 337.922
|
||||
m 359.922 334.32 l 359.922 13.359 m 359.922 16.961 l 369.121 337.922 m
|
||||
369.121 334.32 l 369.121 13.359 m 369.121 16.961 l 378.238 337.922 m 378.238
|
||||
334.32 l 378.238 13.359 m 378.238 16.961 l 387.441 337.922 m 387.441 334.32
|
||||
l 387.441 13.359 m 387.441 16.961 l 396.559 337.922 m 396.559 334.32 l
|
||||
396.559 13.359 m 396.559 16.961 l 405.762 337.922 m 405.762 334.32 l 405.762
|
||||
13.359 m 405.762 16.961 l 405.762 337.922 m 398.559 337.922 l 398.566 337.922
|
||||
l S Q
|
||||
BT
|
||||
12 0 0 12 412.4 42.48 Tm
|
||||
/f-0-0 1 Tf
|
||||
(0)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
405.762 327.121 m 402.16 327.121 l 405.762 316.32 m 402.16 316.32 l 405.762
|
||||
305.441 m 402.16 305.441 l 405.762 294.641 m 402.16 294.641 l 405.762 283.84
|
||||
m 398.559 283.84 l 398.566 283.84 l S Q
|
||||
BT
|
||||
12 0 0 12 412.4 96.56 Tm
|
||||
/f-0-0 1 Tf
|
||||
(50)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
405.762 273.039 m 402.16 273.039 l 405.762 262.16 m 402.16 262.16 l 405.762
|
||||
251.359 m 402.16 251.359 l 405.762 240.559 m 402.16 240.559 l 405.762 229.762
|
||||
m 398.559 229.762 l 398.566 229.762 l S Q
|
||||
BT
|
||||
12 0 0 12 412.4 150.64 Tm
|
||||
/f-0-0 1 Tf
|
||||
(100)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
405.762 218.879 m 402.16 218.879 l 405.762 208.078 m 402.16 208.078 l 405.762
|
||||
197.281 m 402.16 197.281 l 405.762 186.48 m 402.16 186.48 l 405.762 175.602
|
||||
m 398.559 175.602 l 398.566 175.602 l S Q
|
||||
BT
|
||||
12 0 0 12 412.4 204.8 Tm
|
||||
/f-0-0 1 Tf
|
||||
(150)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
405.762 164.801 m 402.16 164.801 l 405.762 154 m 402.16 154 l 405.762 143.199
|
||||
m 402.16 143.199 l 405.762 132.398 m 402.16 132.398 l 405.762 121.52 m
|
||||
398.559 121.52 l 398.566 121.52 l S Q
|
||||
BT
|
||||
12 0 0 12 412.4 258.88 Tm
|
||||
/f-0-0 1 Tf
|
||||
(200)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
405.762 110.719 m 402.16 110.719 l 405.762 99.922 m 402.16 99.922 l 405.762
|
||||
89.121 m 402.16 89.121 l 405.762 78.238 m 402.16 78.238 l 405.762 67.441
|
||||
m 398.559 67.441 l 398.566 67.441 l S Q
|
||||
BT
|
||||
12 0 0 12 412.4 312.96 Tm
|
||||
/f-0-0 1 Tf
|
||||
(250)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
405.762 56.641 m 402.16 56.641 l 405.762 45.84 m 402.16 45.84 l 405.762
|
||||
34.961 m 402.16 34.961 l 405.762 24.16 m 402.16 24.16 l 405.762 13.359
|
||||
m 398.559 13.359 l 398.566 13.359 l S Q
|
||||
BT
|
||||
12 0 0 12 412.4 367.04 Tm
|
||||
/f-0-0 1 Tf
|
||||
(300)Tj
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
57.52 13.359 m 57.52 337.922 l 405.762 337.922 l 405.762 13.359 l h
|
||||
57.52 13.359 m 57.527 13.359 l S Q
|
||||
BT
|
||||
-0.000000000000002204 12 -12 -0.000000000000002204 14.08 163.566094 Tm
|
||||
/f-0-0 1 Tf
|
||||
[(Absorption [mA)32(u])]TJ
|
||||
0.111328 -36.853333 Td
|
||||
[(c\(Imidazol\) [mM])]TJ
|
||||
12 0 0 12 195.998438 6.48 Tm
|
||||
[(V)80(olumen [mL])]TJ
|
||||
ET
|
||||
301.281 240.961 m 303.68 240.961 l 301.281 234.32 l 298.879 240.961 l h
|
||||
f
|
||||
q 1 0 0 -1 0 384 cm
|
||||
301.281 143.039 m 303.68 143.039 l 301.281 149.68 l 298.879 143.039 l 301.281
|
||||
143.039 l 301.281 123.68 m 301.281 143.039 l 301.289 143.039 l S Q
|
||||
347.121 338.32 m 349.52 338.32 l 347.121 331.68 l 344.719 338.32 l h
|
||||
f
|
||||
q 1 0 0 -1 0 384 cm
|
||||
347.121 45.68 m 349.52 45.68 l 347.121 52.32 l 344.719 45.68 l 347.121
|
||||
45.68 l 347.121 26.32 m 347.121 45.68 l 347.129 45.68 l S Q
|
||||
389.281 292.879 m 391.68 292.879 l 389.281 286.238 l 386.879 292.879 l
|
||||
h
|
||||
f
|
||||
q 1 0 0 -1 0 384 cm
|
||||
389.281 91.121 m 391.68 91.121 l 389.281 97.762 l 386.879 91.121 l 389.281
|
||||
91.121 l 389.281 71.762 m 389.281 91.121 l 389.289 91.121 l S Q
|
||||
301.281 240.961 m 303.68 240.961 l 301.281 234.32 l 298.879 240.961 l h
|
||||
f
|
||||
q 1 0 0 -1 0 384 cm
|
||||
301.281 143.039 m 303.68 143.039 l 301.281 149.68 l 298.879 143.039 l 301.281
|
||||
143.039 l 301.281 123.68 m 301.281 143.039 l 301.289 143.039 l S Q
|
||||
347.121 338.32 m 349.52 338.32 l 347.121 331.68 l 344.719 338.32 l h
|
||||
f
|
||||
q 1 0 0 -1 0 384 cm
|
||||
347.121 45.68 m 349.52 45.68 l 347.121 52.32 l 344.719 45.68 l 347.121
|
||||
45.68 l 347.121 26.32 m 347.121 45.68 l 347.129 45.68 l S Q
|
||||
389.281 292.879 m 391.68 292.879 l 389.281 286.238 l 386.879 292.879 l
|
||||
h
|
||||
f
|
||||
q 1 0 0 -1 0 384 cm
|
||||
389.281 91.121 m 391.68 91.121 l 389.281 97.762 l 386.879 91.121 l 389.281
|
||||
91.121 l 389.281 71.762 m 389.281 91.121 l 389.289 91.121 l S Q
|
||||
BT
|
||||
12 0 0 12 223.2775 352.64 Tm
|
||||
/f-0-0 1 Tf
|
||||
[(Absorption)]TJ
|
||||
ET
|
||||
q 1 0 0 -1 0 384 cm
|
||||
284.48 27.762 m 318.238 27.762 l 57.52 305.922 m 58.078 298.559 l 58.641
|
||||
295.922 l 59.121 286.398 l 59.68 289.84 l 60.238 286.32 l 60.801 284.238
|
||||
l 61.359 277.68 l 61.84 276.879 l 62.398 270.961 l 62.961 265.121 l 63.52
|
||||
254.398 l 64 272.801 l 64.559 264.641 l 65.121 256 l 65.68 249.68 l 66.238
|
||||
244.559 l 66.719 238.879 l 67.281 238.48 l 67.84 233.922 l 68.398 233.84
|
||||
l 68.961 237.121 l 69.441 233.281 l 70 234.32 l 70.559 232 l 71.121 229.121
|
||||
l 71.68 229.762 l 72.16 227.281 l 72.719 224.398 l 73.281 215.199 l 73.84
|
||||
187.441 l 74.32 135.441 l 74.879 55.039 l 75.121 13.359 l 141.121 13.359
|
||||
m 141.199 15.121 l 141.68 44.16 l 142.238 70 l 142.801 93.039 l 143.359
|
||||
113.922 l 143.84 132.398 l 144.398 148.48 l 144.961 162.801 l 145.52 175.52
|
||||
l 146.078 186.801 l 146.559 196.559 l 147.121 205.199 l 147.68 213.121
|
||||
l 148.238 220.32 l 148.801 227.039 l 149.281 232.879 l 149.84 238.238 l
|
||||
150.398 243.039 l 150.961 247.52 l 151.52 250.719 l 152 253.52 l 152.559
|
||||
256.078 l 153.121 258.48 l 153.68 260.719 l 154.16 262.879 l 154.719 264.879
|
||||
l 155.281 266.719 l 155.84 268.238 l 156.398 269.68 l 156.879 271.121 l
|
||||
157.441 272.398 l 158 273.762 l 158.559 274.879 l 159.121 275.762 l 159.602
|
||||
276.48 l 160.16 277.121 l 160.719 277.922 l 161.281 278.641 l 161.84 279.441
|
||||
l 162.32 280.32 l 162.879 281.039 l 163.441 281.121 l 164 282.48 l 164.48
|
||||
282.879 l 165.039 283.281 l 165.602 283.441 l 166.16 283.602 l 166.719
|
||||
283.68 l 167.199 283.762 l 167.762 283.84 l 169.922 283.84 l 170.48 283.922
|
||||
l 171.039 284.16 l 171.602 284.48 l 172.16 284.801 l 172.641 285.121 l
|
||||
173.199 285.441 l 173.762 285.68 l 174.32 286.078 l 174.801 286.48 l 175.359
|
||||
286.719 l 175.922 286.879 l 177.039 287.52 l 177.52 287.84 l 178.078 288.238
|
||||
l 178.641 288.32 l 179.199 286.801 l 179.762 288.16 l 180.238 287.922 l
|
||||
180.801 287.762 l 181.359 287.441 l 181.922 286.961 l 182.48 286.48 l 182.961
|
||||
286.238 l 183.52 286.16 l 184.078 286.078 l 184.641 285.84 l 185.121 285.441
|
||||
l 186.238 284.48 l 186.801 284.16 l 187.359 284.32 l 187.84 284.879 l 188.398
|
||||
285.52 l 188.961 286.48 l 189.52 286.961 l 190.078 287.039 l 190.559 287.039
|
||||
l 191.121 286.961 l 192.238 286.961 l 192.801 286.801 l 193.281 286.641
|
||||
l 193.84 286.48 l 194.398 286.398 l 194.961 286.238 l 195.441 286.641 l
|
||||
196 288.961 l 196.559 291.199 l 197.121 293.281 l 197.68 295.121 l 198.16
|
||||
297.039 l 198.719 297.359 l 199.281 299.84 l 199.84 301.441 l 200.398 301.922
|
||||
l 200.879 302.078 l 201.441 302 l 202 301.52 l 202.559 299.84 l 203.121
|
||||
296.801 l 203.602 292.641 l 204.16 287.199 l 204.719 280.238 l 205.281
|
||||
268.719 l 205.762 218.801 l 206.32 164.16 l 206.879 167.121 l 207.441 197.52
|
||||
l 208 219.68 l 208.48 232.48 l 209.039 241.762 l 209.602 249.199 l 210.16
|
||||
255.602 l 210.719 260.641 l 211.199 264.801 l 211.762 267.281 l 212.32
|
||||
268.078 l 212.879 272.641 l 213.441 272.801 l 213.922 273.602 l 214.48 274.398
|
||||
l 215.039 274.801 l 215.602 274.961 l 216.078 276.32 l 216.641 279.52 l
|
||||
217.199 278.559 l 217.762 278.32 l 218.32 279.281 l 218.801 280.398 l 219.359
|
||||
281.281 l 219.922 281.922 l 220.48 282.398 l 221.039 282.879 l 221.52 283.441
|
||||
l 222.078 283.762 l 222.641 283.68 l 223.199 283.359 l 223.762 283.199
|
||||
l 224.238 283.441 l 224.801 284.078 l 225.359 284.801 l 225.922 285.602
|
||||
l 226.398 286.398 l 226.961 287.039 l 227.52 287.68 l 228.078 288.16 l 228.641
|
||||
288.641 l 229.121 289.039 l 229.68 289.281 l 230.238 289.281 l 230.801
|
||||
289.039 l 231.359 288.879 l 231.84 289.039 l 232.398 289.281 l 232.961 289.68
|
||||
l 233.52 290 l 234 290.16 l 234.559 290.16 l 235.121 289.922 l 235.68 289.762
|
||||
l 236.238 289.68 l 236.719 289.68 l 237.281 289.602 l 238.398 288.961 l
|
||||
238.961 288.801 l 239.441 288.879 l 240 289.039 l 240.559 289.359 l 241.121
|
||||
289.762 l 241.68 290 l 242.16 290.238 l 242.719 290.32 l 243.281 290.48
|
||||
l 243.84 290.719 l 244.32 290.879 l 244.879 290.879 l 245.441 290.801 l
|
||||
246 290.719 l 246.559 290.801 l 247.039 291.121 l 247.602 291.52 l 248.16
|
||||
292.078 l 248.719 292.559 l 249.281 293.039 l 249.762 293.359 l 250.32
|
||||
293.602 l 250.879 293.922 l 251.441 294.238 l 252 294.559 l 252.48 294.641
|
||||
l 253.039 294.559 l 253.602 294.398 l 254.16 294.48 l 254.641 294.719 l
|
||||
255.199 295.039 l 255.762 295.441 l 256.32 295.84 l 256.879 296 l 257.359
|
||||
296.16 l 257.922 296.238 l 258.48 296.32 l 259.039 296.48 l 259.602 296.559
|
||||
l 260.078 296.559 l 260.641 296 l 261.199 295.441 l 261.762 295.441 l 262.32
|
||||
295.52 l 262.801 295.762 l 263.359 296 l 263.922 296.238 l 264.48 296.559
|
||||
l 264.961 296.719 l 265.52 296.961 l 266.078 297.359 l 266.641 297.922
|
||||
l 267.199 298.238 l 267.68 298.398 l 268.238 298.398 l 268.801 298.559 l
|
||||
269.359 298.801 l 269.922 298.879 l 270.398 298.961 l 270.961 299.199 l
|
||||
271.52 299.281 l 272.078 298.961 l 272.641 298.48 l 273.121 298.238 l 273.68
|
||||
298.16 l 274.238 298.16 l 274.801 298 l 275.281 297.762 l 275.84 297.68
|
||||
l 276.398 297.68 l 276.961 298 l 277.52 298.559 l 278 299.121 l 278.559
|
||||
299.602 l 279.121 299.84 l 279.68 299.922 l 280.238 299.84 l 280.719 299.68
|
||||
l 281.281 299.68 l 281.84 299.602 l 282.398 299.359 l 282.961 299.039 l
|
||||
283.441 298.641 l 284 298.16 l 284.559 297.922 l 285.121 297.762 l 285.602
|
||||
297.68 l 286.719 297.359 l 287.281 297.199 l 287.84 297.121 l 288.32 297.039
|
||||
l 288.879 297.039 l 289.441 296.801 l 290 296.32 l 290.559 295.68 l 291.039
|
||||
295.121 l 291.602 294.641 l 292.16 294.32 l 292.719 293.602 l 293.281 291.602
|
||||
l 293.762 286.078 l 294.32 272.078 l 294.879 249.199 l 295.441 225.602
|
||||
l 295.922 208.32 l 296.48 197.441 l 297.039 190.801 l 297.602 186.238 l
|
||||
298.16 182.238 l 298.641 178 l 299.199 172.961 l 299.762 167.922 l 300.32
|
||||
164.238 l 300.879 160 l 301.359 157.84 l 301.922 157.359 l 302.48 158.398
|
||||
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1946
img/ergebnisse/cd/cd.eps
Normal file
1946
img/ergebnisse/cd/cd.eps
Normal file
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Load diff
349
img/ergebnisse/cd/cd.svg
Normal file
349
img/ergebnisse/cd/cd.svg
Normal file
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<text><tspan> 190</tspan></text>
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<text><tspan> 230</tspan></text>
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<text><tspan> 240</tspan></text>
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<text><tspan> 250</tspan></text>
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<text><tspan> 260</tspan></text>
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<g style="fill:none; color:black; stroke:currentColor; stroke-width:1.00; stroke-linecap:butt; stroke-linejoin:miter">
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<path d='M63.6,16.7 L63.6,422.4 L575.0,422.4 L575.0,16.7 L63.6,16.7 Z h0.01'/> <g transform="translate(17.6,219.6) rotate(270)" style="stroke:none; fill:black; font-family:Linux Biolinum O; font-size:12.00pt; text-anchor:middle">
|
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<text><tspan>Δε [mdeg M</tspan><tspan font-size="9.6pt" dy="-6.00pt">-1</tspan><tspan font-size="12.0pt" dy="6.00pt">cm</tspan><tspan font-size="9.6pt" dy="-6.00pt">-1</tspan><tspan font-size="12.0pt" dy="6.00pt">]</tspan></text>
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<text><tspan>λ [nm]</tspan></text>
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<g id="gnuplot_plot_1" ><title>gnuplot_plot_1</title>
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<text><tspan>PPDK (experimentell)</tspan></text>
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<text><tspan>Δε [mdeg M</tspan><tspan font-size="9.6pt" dy="-6.00pt">-1</tspan><tspan font-size="12.0pt" dy="6.00pt">cm</tspan><tspan font-size="9.6pt" dy="-6.00pt">-1</tspan><tspan font-size="12.0pt" dy="6.00pt">]</tspan></text>
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4 M q 1 0 0 -1 0 384 cm
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44.238 337.922 m 51.441 337.922 l 460 337.922 m 452.801 337.922 l 452.809
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337.922 l S Q
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BT
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12 0 0 12 32.021875 42.480005 Tm
|
||||
/f-0-0 1 Tf
|
||||
(0)Tj
|
||||
ET
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||||
q 1 0 0 -1 0 384 cm
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||||
44.238 319.922 m 47.84 319.922 l 460 319.922 m 456.398 319.922 l 44.238
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301.84 m 51.441 301.84 l 460 301.84 m 452.801 301.84 l 452.809 301.84 l
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S Q
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BT
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||||
12 0 0 12 32.021875 78.56001 Tm
|
||||
/f-0-0 1 Tf
|
||||
(1)Tj
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||||
ET
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q 1 0 0 -1 0 384 cm
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||||
44.238 283.84 m 47.84 283.84 l 460 283.84 m 456.398 283.84 l 44.238 265.762
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||||
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BT
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12 0 0 12 32.021875 114.63999 Tm
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/f-0-0 1 Tf
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||||
(2)Tj
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ET
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q 1 0 0 -1 0 384 cm
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44.238 247.762 m 47.84 247.762 l 460 247.762 m 456.398 247.762 l 44.238
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/f-0-0 1 Tf
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(3)Tj
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ET
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q 1 0 0 -1 0 384 cm
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BT
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(4)Tj
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ET
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q 1 0 0 -1 0 384 cm
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(5)Tj
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ET
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S Q
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BT
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|
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/f-0-0 1 Tf
|
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(6)Tj
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ET
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44.238 103.52 m 47.84 103.52 l 460 103.52 m 456.398 103.52 l 44.238 85.52
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(7)Tj
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(8)Tj
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44.238 31.359 m 47.84 31.359 l 460 31.359 m 456.398 31.359 l 44.238 13.359
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12 0 0 12 32.021875 367.039999 Tm
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(9)Tj
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ET
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44.238 337.922 m 44.238 330.719 l 44.238 13.359 m 44.238 20.559 l 44.246
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20.559 l S Q
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BT
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12 0 0 12 41.450937 28.080005 Tm
|
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/f-0-0 1 Tf
|
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(0)Tj
|
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ET
|
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q 1 0 0 -1 0 384 cm
|
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64 337.922 m 64 334.32 l 64 13.359 m 64 16.961 l 83.84 337.922 m 83.84
|
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BT
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12 0 0 12 140.410938 28.080005 Tm
|
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/f-0-0 1 Tf
|
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(5)Tj
|
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ET
|
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q 1 0 0 -1 0 384 cm
|
||||
163.039 337.922 m 163.039 334.32 l 163.039 13.359 m 163.039 16.961 l 182.801
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330.719 l 242.238 13.359 m 242.238 20.559 l 242.246 20.559 l S Q
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BT
|
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12 0 0 12 236.661865 28.080005 Tm
|
||||
/f-0-0 1 Tf
|
||||
(10)Tj
|
||||
ET
|
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q 1 0 0 -1 0 384 cm
|
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262 337.922 m 262 334.32 l 262 13.359 m 262 16.961 l 281.84 337.922 m 281.84
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13.359 m 341.199 20.559 l 341.207 20.559 l S Q
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BT
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12 0 0 12 335.621875 28.080005 Tm
|
||||
/f-0-0 1 Tf
|
||||
(15)Tj
|
||||
ET
|
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q 1 0 0 -1 0 384 cm
|
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361.039 337.922 m 361.039 334.32 l 361.039 13.359 m 361.039 16.961 l 380.801
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12 0 0 12 434.661865 28.080005 Tm
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/f-0-0 1 Tf
|
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(20)Tj
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ET
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460 337.922 m 460 334.32 l 460 13.359 m 460 16.961 l 44.238 13.359 m 44.238
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[(Messpu)-3(nkte)]TJ
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1 g
|
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107.199 51.039 m 107.199 49.051 105.59 47.441 103.602 47.441 c 101.613
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0.7992 w
|
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1 g
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1 g
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|
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|
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|
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0.8 w
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|
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1 g
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0 g
|
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0.7992 w
|
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|
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|
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|
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|
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212.559 189.359 c 214.547 189.359 216.16 190.973 216.16 192.961 c h
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|
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0 g
|
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|
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|
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|
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0 g
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S Q
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%%Trailer
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end restore
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149
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|
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|
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|
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|
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|
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|
||||
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|
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|
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|
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|
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|
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|
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|
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|
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|
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|
||||
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|
||||
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|
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|
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|
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|
||||
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|
||||
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|
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|
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|
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|
||||
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|
||||
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|
||||
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|
||||
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|
||||
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|
||||
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694
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(0.4)Tj
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|
||||
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||||
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|
||||
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|
||||
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|
||||
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|
||||
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|
||||
(max)Tj
|
||||
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|
||||
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|
||||
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|
||||
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|
||||
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|
||||
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1 g
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|
||||
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|
||||
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|
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|
||||
0 g
|
||||
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|
||||
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|
||||
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|
||||
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|
||||
S Q
|
||||
1 g
|
||||
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M183.1,422.4 L183.1,417.9 M183.1,16.7 L183.1,21.2 M191.0,422.4 L191.0,417.9 M191.0,16.7 L191.0,21.2
|
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M199.0,422.4 L199.0,417.9 M199.0,16.7 L199.0,21.2 M206.9,422.4 L206.9,417.9 M206.9,16.7 L206.9,21.2
|
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M214.8,422.4 L214.8,417.9 M214.8,16.7 L214.8,21.2 M222.7,422.4 L222.7,417.9 M222.7,16.7 L222.7,21.2
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M230.6,422.4 L230.6,417.9 M230.6,16.7 L230.6,21.2 M238.5,422.4 L238.5,417.9 M238.5,16.7 L238.5,21.2
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M238.5,422.4 L238.5,413.4 M238.5,16.7 L238.5,25.7 h0.01'/> <g transform="translate(238.5,444.9)" style="stroke:none; fill:black; font-family:Linux Biolinum O; font-size:12.00pt; text-anchor:middle">
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M262.3,422.4 L262.3,417.9 M262.3,16.7 L262.3,21.2 M270.2,422.4 L270.2,417.9 M270.2,16.7 L270.2,21.2
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M278.1,422.4 L278.1,417.9 M278.1,16.7 L278.1,21.2 M286.0,422.4 L286.0,417.9 M286.0,16.7 L286.0,21.2
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M294.0,422.4 L294.0,417.9 M294.0,16.7 L294.0,21.2 M301.9,422.4 L301.9,417.9 M301.9,16.7 L301.9,21.2
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M309.8,422.4 L309.8,417.9 M309.8,16.7 L309.8,21.2 M317.7,422.4 L317.7,417.9 M317.7,16.7 L317.7,21.2
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M317.7,422.4 L317.7,413.4 M317.7,16.7 L317.7,25.7 h0.01'/> <g transform="translate(317.7,444.9)" style="stroke:none; fill:black; font-family:Linux Biolinum O; font-size:12.00pt; text-anchor:middle">
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<text><tspan> 0.6</tspan></text>
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M341.5,422.4 L341.5,417.9 M341.5,16.7 L341.5,21.2 M349.4,422.4 L349.4,417.9 M349.4,16.7 L349.4,21.2
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M357.3,422.4 L357.3,417.9 M357.3,16.7 L357.3,21.2 M365.2,422.4 L365.2,417.9 M365.2,16.7 L365.2,21.2
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M373.1,422.4 L373.1,417.9 M373.1,16.7 L373.1,21.2 M381.0,422.4 L381.0,417.9 M381.0,16.7 L381.0,21.2
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M389.0,422.4 L389.0,417.9 M389.0,16.7 L389.0,21.2 M396.9,422.4 L396.9,417.9 M396.9,16.7 L396.9,21.2
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M396.9,422.4 L396.9,413.4 M396.9,16.7 L396.9,25.7 h0.01'/> <g transform="translate(396.9,444.9)" style="stroke:none; fill:black; font-family:Linux Biolinum O; font-size:12.00pt; text-anchor:middle">
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<text><tspan> 0.8</tspan></text>
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M420.6,422.4 L420.6,417.9 M420.6,16.7 L420.6,21.2 M428.5,422.4 L428.5,417.9 M428.5,16.7 L428.5,21.2
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M436.5,422.4 L436.5,417.9 M436.5,16.7 L436.5,21.2 M444.4,422.4 L444.4,417.9 M444.4,16.7 L444.4,21.2
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<text><tspan> 1</tspan></text>
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<text><tspan> 1.2</tspan></text>
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<text><tspan>v</tspan><tspan font-size="9.6pt" dy="3.60pt">0</tspan><tspan font-size="12.0pt" dy="-3.60pt"> [mM min</tspan><tspan font-size="9.6pt" dy="-6.00pt">-1</tspan><tspan font-size="12.0pt" dy="6.00pt">]</tspan></text>
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<g transform="translate(327.6,471.9)" style="stroke:none; fill:black; font-family:Linux Biolinum O; font-size:12.00pt; text-anchor:middle">
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<text><tspan>c(ATP) [mM]</tspan></text>
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<text><tspan>R</tspan><tspan font-size="9.6pt" dy="-6.00pt">2</tspan><tspan font-size="12.0pt" dy="6.00pt"> = 0,906</tspan></text>
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<text><tspan>K</tspan><tspan font-size="9.6pt" dy="3.60pt">M</tspan><tspan font-size="12.0pt" dy="-3.60pt"> = (0,050 ± 0,009) mM</tspan></text>
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<text><tspan>v</tspan><tspan font-size="9.6pt" dy="3.60pt">max</tspan><tspan font-size="12.0pt" dy="-3.60pt"> = (0,095 ± 0,005) mM min</tspan><tspan font-size="9.6pt" dy="-6.00pt">-1</tspan><tspan font-size="12.0pt" dy="6.00pt"></tspan></text>
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<g id="gnuplot_plot_1" ><title>gnuplot_plot_1</title>
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<text><tspan>Messpunkte</tspan></text>
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After Width: | Height: | Size: 13 KiB |
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<text><tspan> 1.2</tspan></text>
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<text><tspan>v</tspan><tspan font-size="9.6pt" dy="3.60pt">0</tspan><tspan font-size="12.0pt" dy="-3.60pt"> [mM min</tspan><tspan font-size="9.6pt" dy="-6.00pt">-1</tspan><tspan font-size="12.0pt" dy="6.00pt">]</tspan></text>
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<text><tspan>K</tspan><tspan font-size="9.6pt" dy="3.60pt">M</tspan><tspan font-size="12.0pt" dy="-3.60pt"> = (0,050 ± 0,009) mM</tspan></text>
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||||
12 0 0 12 453.097656 28.080005 Tm
|
||||
/f-0-0 1 Tf
|
||||
(2.5)Tj
|
||||
ET
|
||||
1 g
|
||||
77.52 125.441 m 77.52 123.453 75.91 121.84 73.922 121.84 c 71.934 121.84
|
||||
70.32 123.453 70.32 125.441 c 70.32 127.43 71.934 129.039 73.922 129.039
|
||||
c 75.91 129.039 77.52 127.43 77.52 125.441 c h
|
||||
f
|
||||
0 g
|
||||
0.7992 w
|
||||
q 1 0 0 -1 0 384 cm
|
||||
77.52 258.559 m 77.52 260.547 75.91 262.16 73.922 262.16 c 71.934 262.16
|
||||
70.32 260.547 70.32 258.559 c 70.32 256.57 71.934 254.961 73.922 254.961
|
||||
c 75.91 254.961 77.52 256.57 77.52 258.559 c h
|
||||
S Q
|
||||
0.8 w
|
||||
q 1 0 0 -1 0 384 cm
|
||||
70.801 13.359 m 70.801 337.922 l 460 337.922 l 460 13.359 l h
|
||||
70.801 13.359 m 70.809 13.359 l S Q
|
||||
BT
|
||||
0 12 -12 0 14.08 173.84812 Tm
|
||||
/f-0-0 1 Tf
|
||||
(v)Tj
|
||||
0 9.6 -9.6 0 17.68 179.402808 Tm
|
||||
(0)Tj
|
||||
0 12 -12 0 14.08 183.865308 Tm
|
||||
([mM min)Tj
|
||||
0 9.6 -9.6 0 8.08 230.787183 Tm
|
||||
[(-1)]TJ
|
||||
0 12 -12 0 14.08 238.455933 Tm
|
||||
(])Tj
|
||||
12 0 0 12 223.73501 6.480005 Tm
|
||||
[(c\(Pyruv)-3(at\) [mM])]TJ
|
||||
6.06875 8.613332 Td
|
||||
(R)Tj
|
||||
9.6 0 0 9.6 303.755322 115.83999 Tm
|
||||
(2)Tj
|
||||
12 0 0 12 308.217822 109.83999 Tm
|
||||
(= 0,947)Tj
|
||||
-1.298153 -1.539998 Td
|
||||
(K)Tj
|
||||
9.6 0 0 9.6 300.186865 87.76001 Tm
|
||||
(M)Tj
|
||||
12 0 0 12 308.699365 91.36001 Tm
|
||||
[(= \(0,270 \261 0,044\))-3( mM)]TJ
|
||||
-1.924947 -1.693335 Td
|
||||
(v)Tj
|
||||
9.6 0 0 9.6 291.154688 67.43999 Tm
|
||||
(max)Tj
|
||||
12 0 0 12 307.879688 71.03999 Tm
|
||||
[(= \(0,093 \261 0,005\))-3( mM min)]TJ
|
||||
9.6 0 0 9.6 432.2625 77.03999 Tm
|
||||
[(-1)]TJ
|
||||
12 0 0 12 337.008447 148.719995 Tm
|
||||
[(Messpu)-3(nkte)]TJ
|
||||
ET
|
||||
1 g
|
||||
269.039 347.602 m 269.039 345.613 267.43 344 265.441 344 c 263.453 344
|
||||
261.84 345.613 261.84 347.602 c 261.84 349.59 263.453 351.199 265.441 351.199
|
||||
c 267.43 351.199 269.039 349.59 269.039 347.602 c h
|
||||
f
|
||||
0 g
|
||||
0.7992 w
|
||||
q 1 0 0 -1 0 384 cm
|
||||
269.039 36.398 m 269.039 38.387 267.43 40 265.441 40 c 263.453 40 261.84
|
||||
38.387 261.84 36.398 c 261.84 34.41 263.453 32.801 265.441 32.801 c 267.43
|
||||
32.801 269.039 34.41 269.039 36.398 c h
|
||||
S Q
|
||||
1 g
|
||||
171.68 336.719 m 171.68 334.73 170.066 333.121 168.078 333.121 c 166.09
|
||||
333.121 164.48 334.73 164.48 336.719 c 164.48 338.707 166.09 340.32 168.078
|
||||
340.32 c 170.066 340.32 171.68 338.707 171.68 336.719 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
171.68 47.281 m 171.68 49.27 170.066 50.879 168.078 50.879 c 166.09 50.879
|
||||
164.48 49.27 164.48 47.281 c 164.48 45.293 166.09 43.68 168.078 43.68 c
|
||||
170.066 43.68 171.68 45.293 171.68 47.281 c h
|
||||
S Q
|
||||
1 g
|
||||
123.121 323.039 m 123.121 321.051 121.508 319.441 119.52 319.441 c 117.531
|
||||
319.441 115.922 321.051 115.922 323.039 c 115.922 325.027 117.531 326.641
|
||||
119.52 326.641 c 121.508 326.641 123.121 325.027 123.121 323.039 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
123.121 60.961 m 123.121 62.949 121.508 64.559 119.52 64.559 c 117.531
|
||||
64.559 115.922 62.949 115.922 60.961 c 115.922 58.973 117.531 57.359 119.52
|
||||
57.359 c 121.508 57.359 123.121 58.973 123.121 60.961 c h
|
||||
S Q
|
||||
1 g
|
||||
98.719 319.121 m 98.719 317.133 97.109 315.52 95.121 315.52 c 93.133 315.52
|
||||
91.52 317.133 91.52 319.121 c 91.52 321.109 93.133 322.719 95.121 322.719
|
||||
c 97.109 322.719 98.719 321.109 98.719 319.121 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
98.719 64.879 m 98.719 66.867 97.109 68.48 95.121 68.48 c 93.133 68.48
|
||||
91.52 66.867 91.52 64.879 c 91.52 62.891 93.133 61.281 95.121 61.281 c 97.109
|
||||
61.281 98.719 62.891 98.719 64.879 c h
|
||||
S Q
|
||||
1 g
|
||||
86.559 208.801 m 86.559 206.812 84.949 205.199 82.961 205.199 c 80.973
|
||||
205.199 79.359 206.812 79.359 208.801 c 79.359 210.789 80.973 212.398 82.961
|
||||
212.398 c 84.949 212.398 86.559 210.789 86.559 208.801 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
86.559 175.199 m 86.559 177.188 84.949 178.801 82.961 178.801 c 80.973
|
||||
178.801 79.359 177.188 79.359 175.199 c 79.359 173.211 80.973 171.602 82.961
|
||||
171.602 c 84.949 171.602 86.559 173.211 86.559 175.199 c h
|
||||
S Q
|
||||
1 g
|
||||
80.48 164.879 m 80.48 162.891 78.867 161.281 76.879 161.281 c 74.891 161.281
|
||||
73.281 162.891 73.281 164.879 c 73.281 166.867 74.891 168.48 76.879 168.48
|
||||
c 78.867 168.48 80.48 166.867 80.48 164.879 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
80.48 219.121 m 80.48 221.109 78.867 222.719 76.879 222.719 c 74.891 222.719
|
||||
73.281 221.109 73.281 219.121 c 73.281 217.133 74.891 215.52 76.879 215.52
|
||||
c 78.867 215.52 80.48 217.133 80.48 219.121 c h
|
||||
S Q
|
||||
q 1 0 0 -1 0 384 cm
|
||||
424.48 231.68 m 424.48 233.668 422.867 235.281 420.879 235.281 c 418.891
|
||||
235.281 417.281 233.668 417.281 231.68 c 417.281 229.691 418.891 228.078
|
||||
420.879 228.078 c 422.867 228.078 424.48 229.691 424.48 231.68 c h
|
||||
S Q
|
||||
1 g
|
||||
269.039 350.48 m 269.039 348.492 267.43 346.879 265.441 346.879 c 263.453
|
||||
346.879 261.84 348.492 261.84 350.48 c 261.84 352.469 263.453 354.078 265.441
|
||||
354.078 c 267.43 354.078 269.039 352.469 269.039 350.48 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
269.039 33.52 m 269.039 35.508 267.43 37.121 265.441 37.121 c 263.453 37.121
|
||||
261.84 35.508 261.84 33.52 c 261.84 31.531 263.453 29.922 265.441 29.922
|
||||
c 267.43 29.922 269.039 31.531 269.039 33.52 c h
|
||||
S Q
|
||||
1 g
|
||||
171.68 335.52 m 171.68 333.531 170.066 331.922 168.078 331.922 c 166.09
|
||||
331.922 164.48 333.531 164.48 335.52 c 164.48 337.508 166.09 339.121 168.078
|
||||
339.121 c 170.066 339.121 171.68 337.508 171.68 335.52 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
171.68 48.48 m 171.68 50.469 170.066 52.078 168.078 52.078 c 166.09 52.078
|
||||
164.48 50.469 164.48 48.48 c 164.48 46.492 166.09 44.879 168.078 44.879
|
||||
c 170.066 44.879 171.68 46.492 171.68 48.48 c h
|
||||
S Q
|
||||
1 g
|
||||
123.121 328.16 m 123.121 326.172 121.508 324.559 119.52 324.559 c 117.531
|
||||
324.559 115.922 326.172 115.922 328.16 c 115.922 330.148 117.531 331.762
|
||||
119.52 331.762 c 121.508 331.762 123.121 330.148 123.121 328.16 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
123.121 55.84 m 123.121 57.828 121.508 59.441 119.52 59.441 c 117.531 59.441
|
||||
115.922 57.828 115.922 55.84 c 115.922 53.852 117.531 52.238 119.52 52.238
|
||||
c 121.508 52.238 123.121 53.852 123.121 55.84 c h
|
||||
S Q
|
||||
1 g
|
||||
98.719 308.32 m 98.719 306.332 97.109 304.719 95.121 304.719 c 93.133 304.719
|
||||
91.52 306.332 91.52 308.32 c 91.52 310.309 93.133 311.922 95.121 311.922
|
||||
c 97.109 311.922 98.719 310.309 98.719 308.32 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
98.719 75.68 m 98.719 77.668 97.109 79.281 95.121 79.281 c 93.133 79.281
|
||||
91.52 77.668 91.52 75.68 c 91.52 73.691 93.133 72.078 95.121 72.078 c 97.109
|
||||
72.078 98.719 73.691 98.719 75.68 c h
|
||||
S Q
|
||||
1 g
|
||||
86.559 190.559 m 86.559 188.57 84.949 186.961 82.961 186.961 c 80.973 186.961
|
||||
79.359 188.57 79.359 190.559 c 79.359 192.547 80.973 194.16 82.961 194.16
|
||||
c 84.949 194.16 86.559 192.547 86.559 190.559 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
86.559 193.441 m 86.559 195.43 84.949 197.039 82.961 197.039 c 80.973 197.039
|
||||
79.359 195.43 79.359 193.441 c 79.359 191.453 80.973 189.84 82.961 189.84
|
||||
c 84.949 189.84 86.559 191.453 86.559 193.441 c h
|
||||
S Q
|
||||
1 g
|
||||
80.48 158.559 m 80.48 156.57 78.867 154.961 76.879 154.961 c 74.891 154.961
|
||||
73.281 156.57 73.281 158.559 c 73.281 160.547 74.891 162.16 76.879 162.16
|
||||
c 78.867 162.16 80.48 160.547 80.48 158.559 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
80.48 225.441 m 80.48 227.43 78.867 229.039 76.879 229.039 c 74.891 229.039
|
||||
73.281 227.43 73.281 225.441 c 73.281 223.453 74.891 221.84 76.879 221.84
|
||||
c 78.867 221.84 80.48 223.453 80.48 225.441 c h
|
||||
S Q
|
||||
1 g
|
||||
269.039 347.84 m 269.039 345.852 267.43 344.238 265.441 344.238 c 263.453
|
||||
344.238 261.84 345.852 261.84 347.84 c 261.84 349.828 263.453 351.441 265.441
|
||||
351.441 c 267.43 351.441 269.039 349.828 269.039 347.84 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
269.039 36.16 m 269.039 38.148 267.43 39.762 265.441 39.762 c 263.453 39.762
|
||||
261.84 38.148 261.84 36.16 c 261.84 34.172 263.453 32.559 265.441 32.559
|
||||
c 267.43 32.559 269.039 34.172 269.039 36.16 c h
|
||||
S Q
|
||||
1 g
|
||||
171.68 342.879 m 171.68 340.891 170.066 339.281 168.078 339.281 c 166.09
|
||||
339.281 164.48 340.891 164.48 342.879 c 164.48 344.867 166.09 346.48 168.078
|
||||
346.48 c 170.066 346.48 171.68 344.867 171.68 342.879 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
171.68 41.121 m 171.68 43.109 170.066 44.719 168.078 44.719 c 166.09 44.719
|
||||
164.48 43.109 164.48 41.121 c 164.48 39.133 166.09 37.52 168.078 37.52
|
||||
c 170.066 37.52 171.68 39.133 171.68 41.121 c h
|
||||
S Q
|
||||
1 g
|
||||
123.121 336.16 m 123.121 334.172 121.508 332.559 119.52 332.559 c 117.531
|
||||
332.559 115.922 334.172 115.922 336.16 c 115.922 338.148 117.531 339.762
|
||||
119.52 339.762 c 121.508 339.762 123.121 338.148 123.121 336.16 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
123.121 47.84 m 123.121 49.828 121.508 51.441 119.52 51.441 c 117.531 51.441
|
||||
115.922 49.828 115.922 47.84 c 115.922 45.852 117.531 44.238 119.52 44.238
|
||||
c 121.508 44.238 123.121 45.852 123.121 47.84 c h
|
||||
S Q
|
||||
1 g
|
||||
98.719 315.762 m 98.719 313.773 97.109 312.16 95.121 312.16 c 93.133 312.16
|
||||
91.52 313.773 91.52 315.762 c 91.52 317.75 93.133 319.359 95.121 319.359
|
||||
c 97.109 319.359 98.719 317.75 98.719 315.762 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
98.719 68.238 m 98.719 70.227 97.109 71.84 95.121 71.84 c 93.133 71.84
|
||||
91.52 70.227 91.52 68.238 c 91.52 66.25 93.133 64.641 95.121 64.641 c 97.109
|
||||
64.641 98.719 66.25 98.719 68.238 c h
|
||||
S Q
|
||||
1 g
|
||||
86.559 203.121 m 86.559 201.133 84.949 199.52 82.961 199.52 c 80.973 199.52
|
||||
79.359 201.133 79.359 203.121 c 79.359 205.109 80.973 206.719 82.961 206.719
|
||||
c 84.949 206.719 86.559 205.109 86.559 203.121 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
86.559 180.879 m 86.559 182.867 84.949 184.48 82.961 184.48 c 80.973 184.48
|
||||
79.359 182.867 79.359 180.879 c 79.359 178.891 80.973 177.281 82.961 177.281
|
||||
c 84.949 177.281 86.559 178.891 86.559 180.879 c h
|
||||
S Q
|
||||
1 g
|
||||
80.48 157.121 m 80.48 155.133 78.867 153.52 76.879 153.52 c 74.891 153.52
|
||||
73.281 155.133 73.281 157.121 c 73.281 159.109 74.891 160.719 76.879 160.719
|
||||
c 78.867 160.719 80.48 159.109 80.48 157.121 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
80.48 226.879 m 80.48 228.867 78.867 230.48 76.879 230.48 c 74.891 230.48
|
||||
73.281 228.867 73.281 226.879 c 73.281 224.891 74.891 223.281 76.879 223.281
|
||||
c 78.867 223.281 80.48 224.891 80.48 226.879 c h
|
||||
S Q
|
||||
BT
|
||||
12 0 0 12 280.055322 134.319995 Tm
|
||||
/f-0-0 1 Tf
|
||||
[(Nichtlinear)7(e Regr)8(ession)]TJ
|
||||
ET
|
||||
1 g
|
||||
77.52 136.719 m 77.52 134.73 75.91 133.121 73.922 133.121 c 71.934 133.121
|
||||
70.32 134.73 70.32 136.719 c 70.32 138.707 71.934 140.32 73.922 140.32
|
||||
c 75.91 140.32 77.52 138.707 77.52 136.719 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
77.52 247.281 m 77.52 249.27 75.91 250.879 73.922 250.879 c 71.934 250.879
|
||||
70.32 249.27 70.32 247.281 c 70.32 245.293 71.934 243.68 73.922 243.68
|
||||
c 75.91 243.68 77.52 245.293 77.52 247.281 c h
|
||||
S Q
|
||||
1 g
|
||||
77.52 114.48 m 77.52 112.492 75.91 110.879 73.922 110.879 c 71.934 110.879
|
||||
70.32 112.492 70.32 114.48 c 70.32 116.469 71.934 118.078 73.922 118.078
|
||||
c 75.91 118.078 77.52 116.469 77.52 114.48 c h
|
||||
f
|
||||
0 g
|
||||
q 1 0 0 -1 0 384 cm
|
||||
77.52 269.52 m 77.52 271.508 75.91 273.121 73.922 273.121 c 71.934 273.121
|
||||
70.32 271.508 70.32 269.52 c 70.32 267.531 71.934 265.922 73.922 265.922
|
||||
c 75.91 265.922 77.52 267.531 77.52 269.52 c h
|
||||
S Q
|
||||
1 0 0 rg
|
||||
0.8 w
|
||||
q 1 0 0 -1 0 384 cm
|
||||
404 246.078 m 437.762 246.078 l 74.719 337.922 m 74.719 191.359 l 78.641
|
||||
148.238 l 82.559 125.039 l 86.559 109.762 l 90.48 98.641 l 94.398 90.16
|
||||
l 98.32 83.441 l 102.238 77.84 l 106.16 73.281 l 110.078 69.281 l 114.078
|
||||
65.922 l 118 62.961 l 121.922 60.398 l 125.84 58.078 l 129.762 56 l 133.68
|
||||
54.16 l 137.602 52.48 l 141.602 50.961 l 145.52 49.52 l 149.441 48.238
|
||||
l 153.359 47.121 l 157.281 46 l 161.199 44.961 l 165.121 44.078 l 169.121
|
||||
43.199 l 173.039 42.32 l 176.961 41.602 l 180.879 40.879 l 184.801 40.238
|
||||
l 188.719 39.602 l 192.641 38.961 l 196.641 38.398 l 200.559 37.84 l 204.48
|
||||
37.359 l 208.398 36.879 l 212.32 36.398 l 216.238 36 l 220.16 35.602 l
|
||||
224.16 35.199 l 228.078 34.801 l 232 34.398 l 235.922 34.078 l 239.84 33.762
|
||||
l 243.762 33.441 l 247.68 33.121 l 251.68 32.801 l 255.602 32.559 l 259.52
|
||||
32.238 l 263.441 32 l 267.359 31.762 l 271.281 31.52 l 275.199 31.281 l
|
||||
279.121 31.039 l 283.121 30.801 l 287.039 30.641 l 290.961 30.398 l 294.879
|
||||
30.238 l 298.801 30 l 302.719 29.84 l 306.641 29.68 l 310.641 29.441 l
|
||||
314.559 29.281 l 318.48 29.121 l 322.398 28.961 l 326.32 28.801 l 330.238
|
||||
28.641 l 334.16 28.559 l 338.16 28.398 l 342.078 28.238 l 346 28.078 l
|
||||
349.922 28 l 353.84 27.84 l 357.762 27.762 l 361.68 27.602 l 365.68 27.52
|
||||
l 369.602 27.359 l 373.52 27.281 l 377.441 27.121 l 381.359 27.039 l 385.281
|
||||
26.961 l 389.199 26.801 l 393.199 26.719 l 397.121 26.641 l 401.039 26.559
|
||||
l 404.961 26.48 l 408.879 26.32 l 412.801 26.238 l 416.719 26.16 l 420.719
|
||||
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440.32 25.68 l 444.238 25.602 l 448.238 25.52 l 452.16 25.441 l 456.078
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25.359 l 460.008 25.359 l S Q
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1 g
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463.602 363.762 m 463.602 361.773 461.988 360.16 460 360.16 c 458.012 360.16
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0 g
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S Q
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456.398 352.09 456.398 354.078 c 456.398 356.066 458.012 357.68 460 357.68
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0 g
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463.602 29.922 m 463.602 31.91 461.988 33.52 460 33.52 c 458.012 33.52
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456.398 31.91 456.398 29.922 c 456.398 27.934 458.012 26.32 460 26.32 c
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S Q
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%%Trailer
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end restore
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%%EOF
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161
img/ergebnisse/kinetik/pyruvat.svg
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161
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After Width: | Height: | Size: 12 KiB |
619
img/ergebnisse/kinetik/pyruvat_editiert.svg
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img/ergebnisse/kinetik/pyruvat_editiert.svg
Normal file
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[Dolphin]
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Timestamp=2012,8,9,19,54,38
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Version=3
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@ -1,8 +1,11 @@
|
|||
\addchap{Abkürzungsverzeichnis}
|
||||
\begin{acronym}[F. trinervia]
|
||||
\begin{acronym}[C. symbiosum]
|
||||
\setlength{\itemsep}{-\parsep}
|
||||
\acro{2YT}{\textit{2x Yeast extract and Tryptone} (engl.)}
|
||||
\acro{3PGAK}{3-Phosphoglycerat Kinase}
|
||||
\acro{AA/BAA}{Acrylamid/Bisacrylamid}
|
||||
\acro{ADP}{Adenosindiphosphat}
|
||||
\acro{AK}{Adenylatkinase}
|
||||
\acro{AMP}{Adenosinmonophosphat}
|
||||
\acro{APS}{Ammoniumperoxodisulfat}
|
||||
\acro{ATP}{Adenosintriphosphat}
|
||||
|
|
@ -29,6 +32,9 @@
|
|||
%\acro{KH2PO4}[\ce{KH2PO4}]{Kaliumdihydrogenphosphat}
|
||||
%\acro{MgCl2}[\ce{MgCl2}]{Magnesiumchlorid}
|
||||
%\acro{MgSO4}[\ce{MgSO4}]{Magnesiumsulfat}
|
||||
\acro{MDH}{Malatdehydrogenase}
|
||||
\acro{ME}{Malatenzym}
|
||||
\acro{MW}{Molekulargewicht}
|
||||
\acro{NADH}{Nicotinamidadenindinukleotid}
|
||||
\acro{NAD-MDH}{NAD-abhängige Malatdehydrogenase}
|
||||
%\acro{NaHCO3}[\ce{NaHCO3}]{Natriumhydrogencarbonat}
|
||||
|
|
@ -37,11 +43,14 @@
|
|||
\acro{p. a.}{\textit{pro analysi}~(lat.)}
|
||||
\acro{PAGE}{Polyacrylamidgelelektrophorese}
|
||||
\acro{PCR}{Polymerase-Kettenreaktion}
|
||||
\acro{PDRP}{PPDK regulierendes Protein}
|
||||
\acro{PEP}{Phosphoenolpyruvat}
|
||||
\acro{PEPCase}{Phosphoenolpyruvat-Carboxylase}
|
||||
\acro{PFK}{Phosphofruktokinase}
|
||||
\acro{PMSF}{Phenylmethylsulfonylfluorid}
|
||||
\acro{PPDK}{Pyruvat-Phosphat Dikinase}
|
||||
\acro{rcf}{\textit{relative centrifugal force}~(engl.)}
|
||||
\acro{RuBisCO}{Ribulose"=1,5"=bisphosphat"=carboxylase/"=oxygenase}
|
||||
\acro{SDS}{Natriumdodecylsulfat}
|
||||
\acro{SLIC}{Sequenz und Ligase unabhängige Klonierung}
|
||||
\acro{SOPMA}{\textit{Self-optimized prediction method}~(engl.)}
|
||||
|
|
|
|||
|
|
@ -1,4 +1,4 @@
|
|||
\selectlanguage{english}
|
||||
\addchap{Abstract}
|
||||
<Abstract>
|
||||
|
||||
\selectlanguage{ngerman}
|
||||
|
|
@ -51,9 +51,28 @@
|
|||
\acs{C. symbiosum}. Sequenzidentitäten sind farbkodiert dargestellt: \SI{100}{\percent} Identität (violett), \SI{75}{\percent} (blau) und
|
||||
\SI{50}{\percent} (rosa).}
|
||||
\shortcaption{Multiples Alignment der Primärstrukturen verschiedener PPDKs}
|
||||
\label{abb:ma}
|
||||
\end{texshade}
|
||||
|
||||
\microtypesetup{protrusion=true}
|
||||
\clearpage
|
||||
|
||||
\section{Evalutation der Homologiemodelle}
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[height=0.5\textheight,keepaspectratio=true]{./img/anhang/FtPPDK_Zm.eps}
|
||||
% FtPPDK_Zm.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=0 -1 474 638
|
||||
\caption{Ramachandran-Plot des von \textit{Zea mays} abgeleiteten Homologiemodells}
|
||||
\label{abb:ramachandran_Zm}
|
||||
\end{figure}
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[height=0.5\textheight,keepaspectratio=true]{./img/anhang/FtPPDK_Cs.eps}
|
||||
% FtPPDK_Cs.svg: 765x990 pixel, 72dpi, 26.99x34.92 cm, bb=0 0 765 990
|
||||
\caption{Ramachandran-Plot des von \acs{C. symbiosum} abgeleiteten Homologiemodells}
|
||||
\label{abb:ramachandran_Cs}
|
||||
\end{figure}
|
||||
|
||||
|
||||
\chapter{Erklärung}
|
||||
Hiermit versichere ich, dass ich die vorliegende Masterarbeit selbstständig verfasst und keine anderen, als die angegebenen Quellen und
|
||||
|
|
|
|||
|
|
@ -1 +1,83 @@
|
|||
\chapter{Diskussion}
|
||||
\chapter{Diskussion}
|
||||
\section{Expression und Reinigung der \acs{PPDK}}
|
||||
Die \acs{PPDK} aus \acs{F. trinervia} konnte erfolgreich in \acs{E. coli} BL21 (DE3) exprimiert werden. Sowohl in einer vierstündigen Expression,
|
||||
als auch in einer Expression über Nacht zeigten sich in der differentiellen Zentrifugation nur geringe unlösliche Anteile, gemessen an der
|
||||
Gesamtmenge Protein (vgl. \ref{sec:ergebnisse_dz}). Dies spricht dafür, dass nur in geringem Maße \textit{Inclusion Bodies} gebildet wurden und legt die
|
||||
Vermutung nahe, das der lösliche Proteinanteil nativ gefaltet ist.
|
||||
|
||||
Dies konnte durch die \acs{CD}-Spektroskopie gestützt werden. Es konnten Sekundärstrukturanteile nachgewiesen werden, welche sich mit
|
||||
Sequenz basierten \textit{ab initio} Vorhersagen, sowie bekannten Strukturdaten decken (vgl. Tabelle \ref{tab:cd}). In einem letzten Schritt konnte die tatsächlich funktionelle
|
||||
Faltung der rekombinanten \acs{PPDK} in einem Aktivitätsassay gezeigt werden.
|
||||
|
||||
Im Rahmen der Reinigung konnten mit steigender Imidazolkonzentration insgesamt drei Fraktionen \acs{PPDK} gewonnen werden, die eine jeweils hohe Reinheit aufweisen (vgl. \ref{abb:reinigung_sds} und sich
|
||||
durch ihre Dispersität unterscheiden (siehe Abbildung \ref{abb:chromatogramm_gefi}). Der Oligomerisierungsgrad beeinflusst offenbar die Bindungsaffinität der \acs{PPDK} an die Säule,
|
||||
was zu einem unterschiedlichen Elutionsverhalten führt. Ursächlich könnte eine zunehmende Maskierung des Hexa-Histidin-Tags in den Oligomeren sein. Eine zuverlässige Größenzuordnung
|
||||
konnte in der Größenausschluschromatographie aufgrund der sehr eng zusammen liegenden Elutionsvolumina nicht vorgenommen werden. Es konnte aber gezeigt werden, dass
|
||||
die bei \SI{250}{\milli\Molar} gewonnene Fraktion monodispers vorliegt und sich somit gut für zukünftige Kristallisationsexperimente eignet.
|
||||
|
||||
Die Proteinausbeute liegt -- betrachtet man nur die monodispers gereinigte Fraktion -- bei etwa \SI{47}{\milli\gram\per\liter} Kulturmedium und ist somit als
|
||||
hoch einzuschätzen. Der in der Literatur beschriebene Effekt der ausgeprägten Kältelabilität der \acs{PPDK} aus \acs{F. trinervia} \citep{Burnell1990},
|
||||
welcher zum Zerfall der funktionellen Tetramere in Monomere führt, ließe sich unter Umständen zur Erhöhung der effektiv nutzbaren Ausbeute einsetzen. Durch eine
|
||||
Vereinigung aller gereinigter PPDK-Fraktionen und anschließender Kältebehandlung sollte zu einer einheitlichen Population der PPDK-Monomere führen. Die Ausbeute
|
||||
beträgt dann etwa \SI{118}{\milli\gram\per\liter} Kultur.
|
||||
|
||||
\section{Aktivität der gereinigten PPDK}
|
||||
Die spezifische Aktivität der \acs{PPDK} aus \acs{F. trinervia} liegt mit \SI[seperr]{0,99(9)}{\Unit\per\milli\gram} in der \acs{PEP}-bildenden Richtung in der Größenordnung
|
||||
von Literaturwerten für \textit{Zea mays} \citep{Hatch1975}. In ähnlicher Weise stimmen auch die bestimmten \ce{K_m}-Werte für ATP \SI[seperr]{50(9)}{\micro\Molar} und
|
||||
Pyruvat \SI[seperr]{270(44)}{\micro\Molar} mit bekannten Literaturwerten von \textit{Zea mays} und \textit{F. brownii} überein \citep{Hatch1975,Ohta1997}, die ebenfalls
|
||||
im zwei- bis dreistelligen mikromolaren Bereich liegen. Ein Vergleich der experimentellen Daten mit Literaturwerten ist in Tabelle \ref{tab:literatur} aufgeführt.
|
||||
%
|
||||
\begin{table}[htb]
|
||||
\centering
|
||||
\begin{threeparttable}
|
||||
\caption[Vergleich der kinetischen Parameter mit Literaturwerten]{Vergleich der kinetischen Parameter mit Literaturwerten der PPDK aus \textit{F. bidentis}, \textit{F. brownii} und \textit{Zea mays}.
|
||||
Die Literaturwerte beziehen sich jeweils auf die \acs{PEP}-bildende Reaktion.}
|
||||
\label{tab:literatur}
|
||||
|
||||
\begin{tabular}{llll}
|
||||
\toprule
|
||||
\multirow{2}{*}{\textbf{Spezies}} & \multicolumn{2}{c}{\textbf{\ce{K_m} [\si{\micro\Molar}]}} & \multirow{2}{*}{\textbf{spez. Aktivität [\si{\Unit\per\milli\gram}]}} \\
|
||||
& \textbf{ATP} & \textbf{Pyruvat} & \\
|
||||
\midrule
|
||||
\acs{F. trinervia} & \num[seperr]{50(9)} & \num[seperr]{270(44)} & \num[seperr]{ 0,99(9)} \\
|
||||
\midrule
|
||||
\textit{F. bidentis} & 25\tnote{2} & 73\tnote{2} & \\
|
||||
\textit{F. brownii} & 88\tnote{2} & 67\tnote{2} & \\
|
||||
\textit{Zea mays} & 95\tnote{2} & 158\tnote{2} & 1,2\tnote{1} \\
|
||||
\bottomrule
|
||||
\end{tabular}
|
||||
\begin{tablenotes}
|
||||
\item[1] \citet{Hatch1975}
|
||||
\item[2] \citet{Ohta1997}
|
||||
\end{tablenotes}
|
||||
|
||||
\end{threeparttable}
|
||||
\end{table}
|
||||
|
||||
\ce{V_{max}} wurde im Rahmen der nichtlinearen Regression für \acs{ATP} und Pyruvat bestimmt. Unter Berücksichtigung der Standardabweichung sind die beiden Werte
|
||||
-- \SI[seperr]{0,095(5)}{\milli\Molar\per\minute} (ATP) und \SI[seperr]{0,093(5)}{\milli\Molar\per\minute} (Pyruvat) -- als identisch anzusehen. Dies entspricht der
|
||||
Erwartung, da die maximale Reaktionsgeschwindigkeit bei Substratsättigung in beiden Fällen identisch sein sollte.
|
||||
|
||||
Der beobachtete Abfalls der gemessenen Reaktionsgeschwindigkeit bei einer \acs{ATP}-Konzentration von \SI{2,5}{\milli\Molar} (Abbildung \ref{abb:kinetik_atp}) ist auf
|
||||
die inhibitorische Wirkung von \acs{ATP}, \acs{ADP} und \acs{AMP} auf die als letzte Stufe des gekoppelten Enzymassays eingesetzte \acs{MDH} zurückzuführen \citep{Harris}. Bei einer \acs{ATP}-Konzentration
|
||||
von \SI{2,5}{\milli\Molar} ist nicht mehr die Pyruvat-Bildung durch die \acs{PPDK}, sondern die Umsetzung des Oxalacetats durch die \acs{MDH} der geschwindigkeitsbestimmende Schritt.
|
||||
|
||||
Zusammenfassend lässt sich festhalten, dass die Aktivität der \acs{PPDK} aus \acs{F. trinervia} in etwa vergleichbar mit anderen bekannten PPDKs ist.
|
||||
|
||||
\section{Homologiemodelle}
|
||||
Die erstellten Homologiemodelle weisen nach einer Analyse mit PROCHECK keine auffälligen Anomalien auf, so dass die Modelle als hinreichend korrekt angesehen werden können.
|
||||
Da die \acs{PPDK} aus \textit{Zea mays} zudem eine hohe Sequenzidentität von \SI{79}{\percent} zur \acs{PPDK} aus \acs{F. trinervia} aufweist, bietet das hierauf basierende
|
||||
Modell eine gute Annäherung an die tatsächliche dreidimensionale Struktur der \acs{PPDK} aus \acs{F. trinervia}, insbesondere in Hinblick auf die räumliche Ausrichtung
|
||||
der Seitenketten in der \acs{PEP}-Bindetasche. In Kombination mit der Identifikation hoch konservierter Sequenzabschnitte können einige für die \acs{PEP}-Bindung und
|
||||
Umsetzung wichtige Reste identifiziert werden \citep{Nakanishi2005}: Arg562, Arg619, Arg669, Glu748, Asp772 und Asn771, sowie Cys834, welches als Protonen-Donor fungiert (Abbildung \ref{abb:substratbindetasche}).
|
||||
Durch gezielte Mutation dieser Reste kann in zukünftigen Experimenten ihr Einfluss auf die Aktivität und Stabilität der \acs{PPDK} untersucht werden.
|
||||
|
||||
\section{Ausblick}
|
||||
Die im Rahmen dieser Arbeit gereinigte \acs{PPDK} liegt in hoher Konzentration, monodispers und hoher Reinheit vor, so dass sie in Kristallisationsexperimente eingesetzt
|
||||
werden kann. Eine erfolgreiche Kristallisation ist dann notwendig für die Strukturaufklärung mittels Röntgenkristallographie. Zusätzlich können anhand der erstellten
|
||||
Homologiemodelle \textit{in silico} Analysen durchgeführt werden. Beispielsweise kann nach Bindetaschen für Effektoren gesucht werden, welche das \textit{Domain-Swiveling}
|
||||
in distinkten Zwischenstadien halten. Sobald Kristallisationsbedingungen für die wildtypische Form etabliert sind, können diese für die Co-Kristallisation mit den gefundenen
|
||||
Effektoren adaptiert werden. Letztere können darüber hinaus direkt mit Hilfe des etablierten Aktivitätsassays auf ihren Einfluss auf die \acs{PPDK}-Aktivität hin untersucht werden.
|
||||
Aus den resultierenden Strukturdaten ließen sich dann intermediäre Konformationen ableiten, die zu einem genaueren Verständnis der Funktionalität
|
||||
der \acs{PPDK} führen können. Mittels einer Netzwerkanalyse ließen sich zusätzlich Regionen innerhalb der \acs{PPDK} identifizieren, welche für die Flexibilität der zentralen
|
||||
Phospho"=Histidin"=Domäne verantwortlich sind. Hier bietet sich ein weiterer Ansatzpunkt um beispielsweise über \textit{Cross-Linking} Zwischenkonformationen zu stabilieren.
|
||||
|
|
@ -1 +1,106 @@
|
|||
\chapter{Einleitung}
|
||||
\chapter{Einleitung}
|
||||
Die \acl{PPDK} (PPDK, EC 2.7.9.1) gehört zur Enzymklasse der Phosphotransferasen (Kinasen) und katalysiert die frei reversible Reaktion:
|
||||
\begin{equation*}
|
||||
\ce{\text{Pyruvat} + \text{ATP} + \text{Pi} <=> \text{PEP} + \text{AMP} + \text{PPi}}
|
||||
\end{equation*}
|
||||
Erstmals beschrieben wurde die \acs{PPDK} von \citet{Slack1968,Reeves1968} in tropischen Gräsern und der parasitären Amöbe
|
||||
\textit{Entamoeba histolytica} \citep{Chastain2011}. Strukturdaten der \acs{PPDK} sind verfügbar aus \textit{Clostridium symbiosum} \citep{Herzberg1996}, \textit{Zea mays} \citep{Nakanishi2005} und \textit{Trypanosomas brucei} \citep{Cosenza2002}.
|
||||
|
||||
\section{Strukturelle Funktionalität der \acs{PPDK}}
|
||||
Die \acs{PPDK} kann in drei Domänen unterteilt werden:
|
||||
\begin{enumerate}
|
||||
\item \acs{PEP}/Pyruvat-Bindedomäne
|
||||
\item Nukleotid-Bindedomäne
|
||||
\item Zentrale Phosphohistidin-Domäne
|
||||
\end{enumerate}
|
||||
|
||||
\begin{figure}[H]
|
||||
\centering
|
||||
\includegraphics[height=0.7\textheight,keepaspectratio=true]{./img/einleitung/ppdk_1vbh.png}
|
||||
% ppdk_1vbh.png: 2480x4004 pixel, 300dpi, 21.00x33.90 cm, bb=0 0 595 961
|
||||
\caption[Struktur der \acs{PPDK} aus \textit{Zea mays}]{Struktur der \acs{PPDK} aus \textit{Zea mays} (PDB: 1VBH). Farblich abgesetzt sind die Nukleotid-Bindedomäne (grün), \acs{PEP}/Pyruvat-Bindedomäne (blau) mit dem Substra \acs{PEP} und einem \ce{Mg2+}-Ion, die Phosphohistidin-Domäne (gelb), sowie die Linker-Peptide(rot). Der katalytische Rest His458 ist durch einen Pfeil hervorgehoben. Die Überlagerung (grau) zeigt die \acs{PPDK} aus \acs{C. symbiosum} (PDB: 1KBL). Das katalytische His455 ist ebenfalls durch einen Pfeil markiert. Es zeigt sich die Torsionsbewegung der zentralen Phosphohistidindomäne.}
|
||||
\label{abb:swivel_domain}
|
||||
\end{figure}
|
||||
|
||||
Verknüpft sind die genannten Domänen über flexible Linker-Peptide. Der Abstand zwischen der Nukleotid-Bindedomäne und der \acs{PEP}/Pyruvat-Bindedomäne beträgt etwa \SI{50}{\angstrom}, so dass eine direkte Interaktion der Substrate mit dem katalytisch
|
||||
aktiven Histidinrest in der zentralen Phosphohistidin-Domäne nicht möglich ist. Daher wurde ein so genannter \textit{Domain-Swiveling}-Mechanismus vorgeschlagen, der eine Torsionsbewegung der Phosphohistidin-Domäne postuliert, wodurch die Übertragung der Phosphatgruppen ermöglicht wird \citep{Herzberg1996,Nakanishi2005}. Das Ausmaß dieser Domänenbewegung ist bei Überlagerung der \acs{PEP}-gebundenen Struktur aus \textit{Zea mays} und der ungebundenen Struktur aus \acs{C. symbiosum} (Abbildung \ref{abb:swivel_domain}).
|
||||
|
||||
\section{Funktion in $\text{C}_\text{4}$-Pflanzen}
|
||||
Am besten untersucht ist die Funktion der \acs{PPDK} in \ce{C_4}"=Pflanzen. In \ce{C_3}-Pflanzen limitiert die Oxygenase-Aktivität der \ac{RuBisCO} die Effizienz
|
||||
der Photosynthese bei warmen Umgebungstemperaturen oder bei Wasserstress. Bei den genannten Bedingungen werden die Spaltöffnungen der Blätter geschlossen um
|
||||
den Wasserverlust durch Verdunstung zu minimieren. Durch das Schließen der Spaltöffnungen wird aber auch die Diffusion von \ce{CO2} in das Blatt vermindert. Die resultierende Absenkung der \ce{CO2}-Konzentration in den photosynthetisch aktiven Zellen führt zu einer Begünstigung der zur \ce{CO2}-Fixierung konkurrierenden Oxygenaseaktivität der \ac{RuBisCO} \citep{Sharkey1989,Cornic2000,Lawlor2002}. Durch die Fixierung von \ce{O2} statt \ce{CO2} wird entsteht als Zwischenprodukt 2-Phosphoglycolat, welches nicht im Calvin-Zyklus verstoffwechselt werden kann und unter zusätzlichem Energieverbrauch über die Photorespiration in 3-Phosphoglycerat umgesetzt wird.
|
||||
|
||||
Die \ce{C_4}"=Kohlenstofffixierung umgeht dieses Problem durch eine räumliche Trennung von \ce{CO2}-Fixierung und Calvin-Zyklus. Hierdurch kann \ce{CO2} konzentriert werden, was der \acs{RuBisCO} eine \ce{CO2} reiche Umgebung zur Verfügung stellt. Hierdurch wird die Oxygenaseaktivität stark reduziert.
|
||||
Die \acs{PPDK} ist hierbei im Stroma der Chloroplasten der Mesophyllzellen lokalisiert und katalysiert dort die Regeneration des primären \ce{CO2}-Akzeptors \ac{PEP} \citep{Hatch2002}.
|
||||
|
||||
\begin{figure}[H]
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/einleitung/C4_photosynthesis_NADP-ME_type.eps}
|
||||
% C4_photosynthesis_NADP-ME_type.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=0 -1 713 233
|
||||
\caption[\ce{C_4}"-Kohlenstofffixierung]{\ce{C_4}"-Kohlenstofffixierung nach dem NADP-\acs{ME} Mechanismus. Dargestellt sind eine Mesophyllzelle (links) und eine Bündelscheidezelle (rechts), sowie die Chloroplasten (grün). \ce{CO2} wird auf \ac{PEP} übertragen, welches in Malat umgewandelt und in die Bündelscheidezelle transportiert wird. Dort wird \ce{CO2} abgespalten und das entstandene Pyruvat zurück in die Chloroplasten der Mesophyllzelle transportiert, wo \ac{PEP} durch die \acs{PPDK} aus Pyruvat regeneriert wird. \citep{Raghavendra2011}}
|
||||
\label{abb:c4_cycle}
|
||||
\end{figure}
|
||||
|
||||
Es existieren unterschiedliche Varianten der \ce{C_4}"=Kohlenstofffixierung, welche sich in drei Punkten voneinander unterscheiden \citep{Raghavendra2011}:
|
||||
\begin{enumerate}
|
||||
\item Dem \ce{C_4}-Körper, der aus den Mesophyllzellen exportiert wird (Malat oder Aspartat)
|
||||
\item Dem \ce{C_3}-Körper, der aus den Bündelscheidezellen zurück in die Mesophyllzellen transportiert wird (Pyruvat oder Alanin)
|
||||
\item Dem zur Decarboxylierung in den Bündelscheidezellen verwendetem Enzym (NADP- bzw. NAD-abhängiges Malatenzym oder \ac{PEP}-Carboxykinase)
|
||||
\end{enumerate}
|
||||
Abbildung \ref{abb:c4_cycle} illustriert die \ce{C_4}"=Kohlenstofffixierung mit Malat als \ce{C_4}-Körper, Pyruvat als \ce{C_3}-Körper und einem NADP-abhängigen Malatenzym, wie sie beispielsweise von Mais oder Zuckerrohr bekannt ist. Allen Varianten gemein ist die zentrale Rolle der \acs{PPDK} bei der Regeneration des primären \ce{CO2}-Akzeptors \ac{PEP}.
|
||||
|
||||
\begin{figure}[H]
|
||||
\centering
|
||||
\includegraphics[width=0.7\textwidth,keepaspectratio=true]{./img/einleitung/aktivierung.png}
|
||||
% aktivierung.png: 716x259 pixel, 72dpi, 25.26x9.14 cm, bb=0 0 716 259
|
||||
\caption[Lichtabhängige Aktivierung der PPDK]{Lichtabhängige Aktivierung der PPDK durch reversible Phosphorylierung eines Threoninrestes \citep{Chastain2011}.}
|
||||
\label{abb:aktivierung_ppdk}
|
||||
\end{figure}
|
||||
|
||||
Die Regulation der \acs{PPDK} erfolgt lichtabhängig durch das \ac{PDRP} über die Phosphorylierung eines Threonin-Restes (vgl. Abbildung \ref{abb:aktivierung_ppdk}) im aktiven Zentrum \citep{Burnell1985,Burnell2006,Chastain2011}.
|
||||
|
||||
\section{Funktion in $\text{C}_\text{3}$-Pflanzen}
|
||||
Über die Funktion der \acs{PPDK} in \ce{C_3}-Pflanzen liegen weitaus weniger gesicherte Erkenntnisse vor. Die \acs{PPDK} kann ubiquitär in Geweben aus \ce{C_3}-Pflanzen nachgewiesen werden, liegt dort aber meist nur in sehr geringen Konzentrationen vor \citep{Chastain2003}. Dies erschwert die biochemische Charakterisierung von \acs{PPDK}s aus \ce{C_3}-Pflanzen \textit{in vivo}, da die Umsetzung der Substrate, sowie die Produktbildung nicht zuverlässig verfolgt werden können. Zudem wird der Substratumsatz durch konkurrierende Reaktionen der Pyruvat-Kinase und \acs{PEP}-Carboxykinase maskiert \citep{Chastain2011}. \textit{Knock-out} Linien von \textit{Arabidopsis thaliana} zeigen keinen veränderten Phänotyp, so dass auch hierüber eine Aussage über die Funktion der \acs{PPDK} in \ce{C_3}-Pflanzen nicht möglich ist \citep{Chastain2011}.
|
||||
|
||||
Untersuchungen an Mais-Samen, in denen die \acs{PPDK} in hohem Maße im Cytoplasma des Endosperms exprimiert wird legen allerdings eine Funktion als ergänzendes Enzym der Glykolyse nahe \citep{Kang2005,Hennen-Bierwagen2009}. Es wurde postuliert, dass die \acs{PPDK} durch die von ihr katalysierte, frei reversible Reaktion den Kohlenstofffluss zwischen verschiedenen Biosynthesewegen
|
||||
ausgleicht \citep{Hennen-Bierwagen2009} und \acs{ATP} in hypoxischen Bereichen des Endosperms zur Verfügung stellt \citep{Chastain2006}.
|
||||
|
||||
\section{Funktion in nicht-pflanzlichen Organismen}
|
||||
|
||||
Die \acs{PPDK} wurde in einigen Archäen, Protozoen und Baterien gefunden, welche in sauerstoffarmen Habitaten leben. Die \acs{PPDK} hat in diesen Organismen die Rolle eines primären oder sekundären Enzyms der Glykolyse. Die freie Reversibilität der katalysierten
|
||||
Reaktion erlaubt sowohl die Synthese von \acs{ATP}, als auch die Bildung von \ac{PEP} im Rahmen der Gluconeogenese \citep{Chastain2011}.
|
||||
%
|
||||
\begin{figure}[H]
|
||||
\centering
|
||||
\includegraphics[width=0.6\textwidth,keepaspectratio=true]{./img/einleitung/ppdk_glycolysis.eps}
|
||||
% ppdk_glycolysis.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=0 -1 536 475
|
||||
\caption[Einfluss der \acs{PPDK} auf die glykolytische ATP-Ausbeute]{Einfluss der \acs{PPDK} auf die glykolytische ATP-Ausbeute. Durch das Zusammenspiel zwischen \ac{AK} und \ac{PPDK} können im Vergleich zur konventionellen Glykolyse drei zusätzliche Moleküle \acs{ATP} je Glukosemolekül gewonnen werden.}
|
||||
\label{abb:glykolyse}
|
||||
\end{figure}
|
||||
%
|
||||
Durch synergetische Effekte zwischen \acl{AK} und \ac{PPDK} kann die \acs{ATP}-Ausbeute von drei Molekülen \acs{ATP} je Molekül Glukose auf fünf Moleküle \acs{ATP} erhöht werden (Abbildung \ref{abb:glykolyse}). Hierbei kann die Bindungsenergie von Pyrophosphat durch die \acs{PPDK} zum Aufbau von \acs{ATP} aus \acs{AMP} genutzt werden \citep{Huang2008}. Die Reaktion läuft dabei in die Pyruvat-formende Richtung ab.
|
||||
|
||||
\section{Kälteinaktivierung}
|
||||
|
||||
Eine besondere Eigenschaft der \acs{PPDK} ist die Kälteinaktivierung. Die funktionelle Form ist ein Tetramer, welches bei Temperaturen unterhalb von \SI{10}{\celsius} in inaktive Di- und Monomere zerfällt \citep{Slack1968,Shirahashi1978,Burnell1990}.
|
||||
Bei der experimentellen Arbeit mit der \acs{PPDK} ergibt sich hieraus die Problematik, dass alle Arbeitsschritte bei Raumtemperatur
|
||||
durchgeführt werden müssen, was aber die Anfälligkeit für die Degradierung durch im Zelllysat enthaltene Proteasen erhöht. Durch Erwärmen auf \SI{30}{\celsius} ist allerdings eine weitgehende Reaktivierung zu erreichen \citep{Burnell1985}.
|
||||
|
||||
Die Kältesensitivität ist zwischen verschiedenen Spezies unterschiedlich stark ausgeprägt. Innerhalb der Gattung \textit{Flaveria} weist die \acs{PPDK} aus \acs{F. trinervia} die höchste Sensitivität auf (vgl. Abbildung \ref{abb:cold_lability}): Nach einer \SI{30}{\minute} Inkubation bei \SI{0}{\celsius} konnte eine Restaktivität von lediglich \SI{10}{\percent} beobachtet werden. Die \acs{PPDK} aus \acs{F. brownii} wies nach der Kältebehandlung eine Restaktivität von \SI{80}{\percent} auf \citep{Burnell1990}.
|
||||
%
|
||||
\begin{figure}[H]
|
||||
\centering
|
||||
\includegraphics[width=0.6\textwidth,keepaspectratio=true]{./img/einleitung/cold_lability.png}
|
||||
% cold_lability.png: 470x467 pixel, 96dpi, 12.43x12.35 cm, bb=0 0 352 350
|
||||
\caption[Kälteinaktivierung der \acs{PPDK}]{Kälteinaktivierung der \acs{PPDK}. Aufgetragen ist die Restaktivität in Abhängigkeit von der Inkubationsdauer bei \SI{0}{\celsius}. Die Restaktivität der \acs{PPDK} aus \acs{F. trinervia} beträgt nach \SI{30}{\minute} lediglich \SI{10}{\percent} \citep{Burnell1990}.}
|
||||
\label{abb:cold_lability}
|
||||
\end{figure}
|
||||
%
|
||||
Durch Sequenzanalysen konnten gezeigt werden, dass nur wenige Aminosäurereste die Kältesensitivität stark beeinflussen \citep{Ohta1997}. Darüberhinaus kann durch Zusatz von Glycerin und reduzierenden Agenzien wie \acs{DTT} die Aktivität zu einem Großteil erhalten bleiben \citep{Shirahashi1978}.
|
||||
|
||||
\section{Zielsetzung}
|
||||
|
||||
Die \acs{PPDK} aus \textit{Zea mays} wurde bereits heterolog in \acs{E. coli} exprimiert \citep{Chastain1996,Nakanishi2003} und gereinigt. Ebenfalls etabliert ist ein photometrischer Aktivitätsassay über die \acs{PEP}-Carboxylase und die Malatdehydrogenase \citep{Jenkins1985,Salahas1990}.
|
||||
|
||||
Auf diesen Ergebnissen aufbauend war es Ziel dieser Arbeit, ein Expressions- und Reinigungsprotokoll für die \acs{PPDK} aus \acs{F. trinervia} zu etablieren, sowie nachzuweisen, dass das gereinigte Enzym funktionell gefalten ist. Darüber hinaus sollten Homologiemodelle erstellt werden, um im Vorfeld zukünftiger Experimente die Identifikation funktionell und strukturell bedeutender Reste zu ermöglichen.
|
||||
|
||||
|
|
@ -5,8 +5,9 @@
|
|||
\label{sec:erg_klonierung}
|
||||
%
|
||||
Sie \acs{SLIC}-Klonierung erfolgte wie in \ref{sec:slic} beschrieben. Die mittels \acs{PCR} synthetisierten \textit{Inserts}
|
||||
(vgl.~\ref{sec:inserts_slic}), sowie der linearisierte Vektor (vgl.~\ref{sec:restriktionsverdau_dna}) wurden auf ein zur Reinigung auf
|
||||
Agarosegel aufgetragen (vgl. \ref{sec:agarose_gelelektrophorese}). Es ergab sich das in Abbildung \ref{abb:agarosegel_vektor_insert} gezeigte Auftrennungsmuster.
|
||||
(vgl.~\ref{sec:inserts_slic}), sowie der linearisierte Vektor (vgl.~\ref{sec:restriktionsverdau_dna}) wurden zur Reinigung auf ein
|
||||
Agarosegel aufgetragen (vgl. \ref{sec:agarose_gelelektrophorese}). Es ergab sich das in Abbildung \ref{abb:agarosegel_vektor_insert} gezeigte
|
||||
Auftrennungsmuster.
|
||||
%
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
|
|
@ -35,7 +36,7 @@ Agarosegel aufgetragen (vgl. \ref{sec:agarose_gelelektrophorese}). Es ergab sich
|
|||
\label{abb:agarosegel_vektor_insert}
|
||||
\end{figure}
|
||||
|
||||
Sowohl Vektor als auch Insert lagen innerhalb der erwarteten Größenordnung (\SI{5720}{\bp} bzw. \SI{2691}{\bp}) auf, so dass beide wie in \ref{sec:slic}
|
||||
Sowohl Vektor als auch Insert lagen innerhalb der erwarteten Größenordnung (\SI{5720}{\bp} bzw. \SI{2691}{\bp}), so dass beide wie in \ref{sec:slic}
|
||||
beschrieben in den Ligationsansatz gegeben wurden. Zur Amplifikation des Vektors wurden dann \acs{E. coli} Zellen des Stamms XL1-Blue mit diesem transformiert
|
||||
(vgl. \ref{sec:transformation}) und auf Agarplatten angezogen. Mit einer Kolonie-PCR (vgl. \ref{sec:kolonie_pcr}) wurde überprüft, ob der untersuchte Klon
|
||||
das korrekte Insert trägt. Das enstsprechende Gelbild ist in Abbildung \ref{abb:kolonie_pcr} dargestellt.
|
||||
|
|
@ -49,9 +50,9 @@ das korrekte Insert trägt. Das enstsprechende Gelbild ist in Abbildung \ref{abb
|
|||
\label{abb:kolonie_pcr}
|
||||
\end{figure}
|
||||
|
||||
Von untersuchten 24 Klonen wiesen fünf ein Plasmid auf, das der erwarteten Größe von \SI{8351}{\bp} (Vektor~+~Insert) entspricht. Die fünf positiven Klone
|
||||
wurden in \SI{5}{\milli\liter} Kulturrörchen über Nacht angezogen und die Plasmid-DNA mit dem QIAprep Spin Miniprep Kit nach Herstellerangaben extrahiert.
|
||||
Proben der isolierten Plasmid-DNA wurden zur Sequenzierung eingeschickt (StarSEQ GmbH, Mainz). Es konnte gezeigt werden, dass das aus Klon \#22 isolierte
|
||||
Von untersuchten 24 Klonen wiesen fünf ein Plasmid auf, welches ein Insert der erwarteten Größe von \SI{2691}{\bp} trägt. Die fünf positiven Klone
|
||||
wurden in \SI{5}{\milli\liter} Kulturrörchen über Nacht angezogen und die Plasmid-DNA mit dem \textit{QIAprep Spin Miniprep Kit} nach Herstellerangaben extrahiert.
|
||||
Proben der isolierten Plasmid-\acs{DNA} wurden zur Sequenzierung eingeschickt (StarSEQ GmbH, Mainz). Es konnte gezeigt werden, dass das aus Klon \#22 isolierte
|
||||
Plasmid im für die \acs{PPDK} kodierenden Bereich keine Mutation trägt. Eine Plasmidkarte von pETEV-16b-ppdk ist in Abbildung \ref{abb:plasmidkarte}
|
||||
aufgeführt.
|
||||
%
|
||||
|
|
@ -59,6 +60,325 @@ aufgeführt.
|
|||
\centering
|
||||
\includegraphics[width=0.8\textwidth,keepaspectratio=true]{./img/ergebnisse/pETEV-16b-ppdk.pdf}
|
||||
% pETEV-16b-ppdk.pdf: 611x480 pixel, 72dpi, 21.55x16.93 cm, bb=0 0 611 480
|
||||
\caption{Plasmidkarte von pETEV-16b-ppdk}
|
||||
\caption[Plasmidkarte von pETEV-16b-ppdk]{Plasmidkarte von pETEV-16b-ppdk. Der für die \acs{PPDK} kodierende Bereich liegt zwischen Position 5392 und 8022.
|
||||
\textit{Upstream} befinden sich die kodierenden Sequenzen für die \acs{TEV}-Schnittstelle und den Hexa-Histidin-Tag. Die Plasmidkarte wurde mit
|
||||
\textit{PlasMapper} \citep{Dong2004} erstellt.}
|
||||
\label{abb:plasmidkarte}
|
||||
\end{figure}
|
||||
|
||||
\section{Expressionsstudien}
|
||||
\label{sec:Expressionsstudien}
|
||||
Die Expressionsstudien (vgl. \ref{sec:expression_ecoli_studien}) zeigten eine deutliche Überexpression der \acs{PPDK} in \acs{E. coli} BL21 (DE3) bei einer Temperatur von \SI{30}{\celsius} und
|
||||
einer \acs{IPTG}-Konzentration von \SI{0,1}{\milli\Molar} (vgl. Abbildung \ref{abb:expressionsstudie}). Der direkte Vergleich zwischen den beiden verwendeten
|
||||
Expressionsstämmen BL21 (DE3) und BL21 Gold (DE3) (Abbildung \ref{abb:expressionsstudie_klein}) zeigt eine deutlich stärkere Überexpression in BL21 (DE3) bei
|
||||
gleichzeitig geringerer IPTG-Konzentration. Eine Expression über Nacht führte zu einer deutlich höheren Proteinausbeute im Vergleich zu einer Zellernte nach
|
||||
\SI{4}{\hour}. Die Identifizierung der \acs{PPDK} erfolgte durch einen Westernblot (vgl. \ref{sec:westernblot}).
|
||||
%
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/sds_gele/Expressionsstudie_19042012_gross.png}
|
||||
% Expressionsstudie_19042012_gross.png: 2480x1828 pixel, 300dpi, 21.00x15.48 cm, bb=0 0 595 439
|
||||
\caption[\acs{SDS}-\acs{PAGE} der Expressionsstudien]{\acs{SDS}-\acs{PAGE} der Expressionsstudien. Aufgetragen wurden Proben der beiden Expressionsstämme
|
||||
BL21 (DE3) und BL21 Gold (DE3) bei Induktion mit unterschiedlicher \acs{IPTG}-Konzentration und einer Temperatur von jeweils \SI{30}{\celsius}.
|
||||
Angegeben sind die Zeitpunkte der Probenentnahme vom Zeitpunkt der Induktion an. Der Größenbereich der \acs{PPDK} ist rot hervorgehoben.}
|
||||
\label{abb:expressionsstudie}
|
||||
\end{figure}
|
||||
%
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\begin{subfigure}[b]{0.4\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/sds_gele/Expressionsstudie_19042012_klein.png}
|
||||
% Expressionsstudie_19042012_klein.png: 2480x2894 pixel, 241dpi, 26.13x30.49 cm, bb=0 0 741 864
|
||||
\caption{}
|
||||
\label{abb:expressionsstudie_klein_sds}
|
||||
\end{subfigure}
|
||||
%
|
||||
~
|
||||
%
|
||||
\begin{subfigure}[b]{0.4\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/sds_gele/Expressionsstudie_19042012_klein_blot.png}
|
||||
% Expressionsstudie_19042012_klein_blot.png: 2480x2894 pixel, 241dpi, 26.13x30.49 cm, bb=0 0 741 864
|
||||
\caption{}
|
||||
\label{abb:expressionsstudie_klein_blot}
|
||||
\end{subfigure}
|
||||
\caption[Vergleich der Expression in BL21 (DE3) und BL21 Gold (DE3)]{Vergleich der Expression in BL21 (DE3) und BL21 Gold (DE3). SDS-PAGE (a) und Westernblot (b). Aufgetragen wurden
|
||||
Expressionsproben nach \SI{0}{\hour} und \SI{4}{\hour} nach Induktion, sowie über Nacht. Verglichen wurden die Expressionsreihen mit dem jeweils besten
|
||||
Expressionsergebnis in BL21 (DE3) bzw. BL21 Gold (DE3). Die Induktion erfolgte mit \SI{0,1}{\milli\Molar} bzw. \SI{1}{\milli\Molar} IPTG.}
|
||||
\label{abb:expressionsstudie_klein}
|
||||
\end{figure}
|
||||
|
||||
\section{Differentielle Zentrifugation}
|
||||
\label{sec:ergebnisse_dz}
|
||||
|
||||
Im Rahmen der Expressionsstudien wurden Zellen von \acs{E. coli} BL21 (DE3) nach \SI{4}{\hour} bzw. nach einer Übernachtexpression geerntet, aufgeschlossen
|
||||
und einer differentiellen Zentrifugation unterzogen (vgl. \ref{sec:differentielle_zentrifugation}). Die Proben wurden mittels eines Westernblots ausgewertet
|
||||
(Abbildung \ref{abb:dz}). Hierbei zeigte sich, dass der überwiegende Anteil der \acs{PPDK} in den Überstandsfraktionen lokalisiert und somit löslich ist.
|
||||
Nur ein geringer Anteil befindet sich in den Pellets nach der Zentrifugation.
|
||||
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\begin{subfigure}[b]{0.4\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/sds_gele/03052012_4h_2min.png}
|
||||
% 03052012_4h_2min.png: 2480x2254 pixel, 300dpi, 21.00x19.08 cm, bb=0 0 595 541
|
||||
\caption{}
|
||||
\label{abb:dz_4h}
|
||||
\end{subfigure}
|
||||
%
|
||||
~
|
||||
%
|
||||
\begin{subfigure}[b]{0.4\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/sds_gele/03052012_üN_2min.png}
|
||||
% 03052012_üN_2min.png: 2480x2141 pixel, 300dpi, 21.00x18.13 cm, bb=0 0 595 514
|
||||
\caption{}
|
||||
\label{abb:dz_on}
|
||||
\end{subfigure}
|
||||
\caption[Westernblots der differentiellen Zentrifugation]{Westernblots der differentiellen Zentrifugation. Die Zellen wurden nach \SI{4}{\hour} (a) bzw. nach
|
||||
einer Übernachtexpression (b) geerntet. Aufgetragen wurden \SI{3}{\micro\liter} Marker (M), Probe nach Zellaufschluss (A), sowie Pellets (P) und Überstände (Ü) der
|
||||
Zentrifugationsschritte (jeweils \SI{10}{\micro\liter}).}
|
||||
\label{abb:dz}
|
||||
\end{figure}
|
||||
|
||||
\section{Native Reinigung der \acs{PPDK}}
|
||||
\label{sec:ergebnisse_reinigung}
|
||||
|
||||
Die Reinigung der \acs{PPDK} erfolgte wie in \ref{sec:affinitaetschromatographie} beschrieben. Im Verlauf der Reinigung konnten vier Proteinfraktionen bei Elution
|
||||
mit \SIlist{50;150;200;250}{\milli\Molar} Imidazol identifiziert werden (vgl. Abbildung \ref{abb:chromatogramm_aff}). In einer nachfolgend durchgeführten
|
||||
\acs{SDS}-\acs{PAGE} mit Westernblot zeigte sich, dass drei Fraktionen (\SIlist{150;200;250}{\milli\Molar}) \acs{PPDK} in hoher Reinheit enthielten.
|
||||
%
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[width=0.9\textwidth,keepaspectratio=true]{img/ergebnisse/aekta/20062012.eps}
|
||||
\caption[Chromatogramm der \acs{PPDK}-Reinigung mittels \acs{IMAC}]{Chromatogramm der \acs{PPDK}-Reinigung mittels \acs{IMAC}. Dargestellt sind die Absorption bei
|
||||
\SI{280}{\nano\meter} (blau) und die Imidazolkonzentration (rot). Die gesammelten \acs{PPDK}-Fraktionen sind durch Pfeile markiert.}
|
||||
\label{abb:chromatogramm_aff}
|
||||
\end{figure}
|
||||
%
|
||||
Die gesammelte PPDK-Fraktionen bei Elution mit \SI{250}{\milli\Molar} Imidazol ließ sich bis auf \SI{37,54(134)}{\milli\gram\per\milli\liter} konzentrieren.
|
||||
In der Expression konnten aus \SI{2}{\liter} Kultur \SI{20}{\gram} Zellen geerntet werden. Da im Zuge der Reinigung \SI{4}{\gram} Zellen aufgeschlossen wurden, ergibt sich eine Ausbeute von etwa
|
||||
\SI{47}{\milli\gram\per\liter}~Kultur (vgl. Tabelle \ref{tab:ausbeute}).
|
||||
%
|
||||
\begin{table}[htb]
|
||||
\centering
|
||||
\caption[\acs{PPDK}-Konzentration und Ausbeute nach Reinigung]{\acs{PPDK}-Konzentration und Ausbeute nach Reinigung. Die Konzentrationen beziehen sich auf den erreichten
|
||||
Wert nach Einengen der fraglichen Fraktionen auf ein Volumen von \SI{500}{\micro\liter}.}
|
||||
\label{tab:ausbeute}
|
||||
\begin{tabular}{lS[seperr,table-format=2.3(3)]S[seperr,table-format=2.3(3)]S[seperr,table-format=2.3(3)]}
|
||||
\toprule
|
||||
\textbf{Fraktion} & \multicolumn{1}{c}{\textbf{c(PPDK) [\si{\milli\gram\per\milli\liter}]}} & \multicolumn{1}{c}{\textbf{m(PPDK) [\si{\milli\gram}]}} & \multicolumn{1}{c}{$\frac{\textbf{m(PPDK)}}{\textbf{V(Kultur)}}$ \textbf{[\si{\milli\gram\per\liter}]}}\\
|
||||
\midrule
|
||||
1 (\SI{150}{\milli\Molar}) & 8,078(1812) & 4,039(906) & 10,098(2265)\\
|
||||
2 (\SI{200}{\milli\Molar}) & 48,935(2556) & 24,466(1278) & 61,165(3195)\\
|
||||
3 (\SI{250}{\milli\Molar}) & 37,536(1334) & 18,768(667) & 46,920(1693)\\
|
||||
\bottomrule
|
||||
\end{tabular}
|
||||
\end{table}
|
||||
%
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\begin{subfigure}[b]{0.4\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/sds_gele/Reinigung_02072012.png}
|
||||
% Reinigung_02072012.png: 2480x2338 pixel, 300dpi, 21.00x19.80 cm, bb=0 0 595 561
|
||||
\caption{}
|
||||
\label{abb:reinigung_sds_a}
|
||||
\end{subfigure}
|
||||
%
|
||||
~
|
||||
%
|
||||
\begin{subfigure}[b]{0.4\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/sds_gele/Reinigung_02072012_blot.png}
|
||||
% Reinigung_02072012.png: 2480x2338 pixel, 300dpi, 21.00x19.80 cm, bb=0 0 595 561
|
||||
\caption{}
|
||||
\label{abb:reinigung_sds_b}
|
||||
\end{subfigure}
|
||||
\caption[\acs{SDS}-\acs{PAGE} und Westernblot der nativen Reinigung]{\acs{SDS}-\acs{PAGE} (a) und Westernblot (b) der nativen Reinigung. Aufgetragen wurden \SI{3}{\micro\liter} Marker (M), \SI{1}{\micro\liter}
|
||||
der Proben des Zelllysates (A), sowie der Zentrifugationen bei \SI{10000}{\xg} (10kÜ) bzw \SI{100000}{\xg} und \SI{10}{\micro\liter} des Durchflusses (D) und nachfolgenden Elutionsfraktionen.
|
||||
Die \acs{PPDK}-Banden sind rot hervorgehoben (MW: \SI{98}{\kilo\dalton})}
|
||||
\label{abb:reinigung_sds}
|
||||
\end{figure}
|
||||
%
|
||||
Zur Bestimmung der Dispersität der gereinigten \acs{PPDK} wurden Proben aller drei Fraktionen mittels einer Größenausschlusschromatographie (vgl. \ref{sec:groessenausschluss}) analysiert.
|
||||
Hierbei zeigte sich, dass lediglich die mit \SI{250}{\milli\Molar} Imidazol eluierte Fraktion monodispers ist, während die mit \SIlist{150;200}{\milli\Molar} eluierten
|
||||
\acs{PPDK}-Fraktionen einen diversen Oligomerisierungsgrad aufwiesen (vgl. Abbildung \ref{abb:chromatogramm_gefi}).
|
||||
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[width=0.8\textwidth,keepaspectratio=true]{./img/ergebnisse/aekta/11072012.eps}
|
||||
% 11072012.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=5 4 461 375
|
||||
\caption[Chromatogramm der Größenausschlusschromatographie]{Chromatogramm der Größenausschlusschromatographie. Die Kurvenverläufe wurden auf die jeweilige Konzentration normalisiert. Fraktion 3 zeigt ein monodisperses Profil,
|
||||
während die \acs{PPDK} in den Fraktionen 1 und 2 in verschiedenen Oligomerisierungszuständen vorliegt. Vertikale Linien markieren die Position der \textit{Peaks}.}
|
||||
\label{abb:chromatogramm_gefi}
|
||||
\end{figure}
|
||||
\clearpage
|
||||
|
||||
\section{\acs{CD}-Spektroskopie}
|
||||
\label{sec:ergebnisse_cd}
|
||||
Das \acs{CD}-Spektrum wurde wie in \ref{sec:cd_spektroskopie} beschrieben aufgenommen. Abbildung \ref{abb:cd_spektrum} zeigt das Spektrum nach Abzug des Pufferspektrums. Deutlich zu erkennen ist
|
||||
ein negativer Cotton-Effekt im Bereich um \SI{210}{\nano\meter}, welcher auf \textalpha-helikale Strukturen hindeutet.
|
||||
Anhand des experimentellen Spektrums wurde eine Sekundärstrukturanalyse mit den Programmen SELCON3 \citep{Sreerama1993}, CONTINLL \citep{Provencher1981}, CDSSTR \citep{Johnson1999} und K2D3 \citep{Louis-Jeune2011} durchgeführt.
|
||||
Die daraus resultierenden, vorhergesagten Spektren sind ebenfalls in Abbildung \ref{abb:cd_spektrum} dargestellt. Alle verwendeten Algorithmen sagen das Spektrum qualitativ
|
||||
korrekt vorher, die geringsten Abweichungen vom experimentellen Spektrum ergeben sich für CONTINLL und CDSSTR. Für die \acs{PPDK} werden in allen Fällen hohe \textalpha"=helikale
|
||||
Strukturanteile vorhergesagt, im Fall von CONTINLL jedoch nur ein sehr geringer Anteil an \textbeta"=Strands (vgl. Tabelle \ref{tab:cd}). Die \textit{ab initio} Vorhersage mit SOPMA,
|
||||
sowie die Ergebnisse von K2D3 und CDSSTR sind weitestgehend deckungsgleich mit den Strukturanteilen der PPDK aus \textit{Zea mays} (Tabelle \ref{tab:cd}).
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[width=0.8\textwidth,keepaspectratio=true]{./img/ergebnisse/cd/cd.eps}
|
||||
% cd.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=2 4 468 375
|
||||
\caption[\acs{CD}-Spektrum von gereinigter PPDK]{\acs{CD}-Spektrum von gereinigter PPDK. Die \acs{PPDK} lag in einer Konzentration von \SI{0,1}{\milli\gram\per\milli\liter} in \SI{50}{\milli\Molar} Kaliumphosphat"=Puffer mit
|
||||
\SI{10}{\milli\Molar} Magnesiumsulfat vor. Es wurden 15 Spektren mit einer Auflösung von \SI{1}{\nano\meter} akkumuliert. Im Bereich um \SI{210}{\nano\meter} zeigt sich ein negativer
|
||||
Cotton-Effekt, welcher charakteristisch für \textalpha-helikale Strukturanteile ist. Farbig dargestellt sind die mit unterschiedlichen Programmen vorhergesagten Spektren.}
|
||||
\label{abb:cd_spektrum}
|
||||
\end{figure}
|
||||
|
||||
\begin{table}[htb]
|
||||
\caption[Vorhersage der Sekundärstrukturanteile]{Vorhersage der Sekundärstrukturanteile. Aufgeführt sind die vorhergesagten Strukturelemente anhand des \acs{CD}-Spektrums. Zum Vergleich dienen die Ergebnisse der sequenzbasierten \textit{ab initio} Vorhersage (SOPMA),
|
||||
sowie die Sekundärstrukturanteile aus der Kristallstruktur der \acs{PPDK} aus \textit{Zea mays} (PDB: 1VBG).}
|
||||
\label{tab:cd}
|
||||
\begin{tabular}{lrrrr}
|
||||
\toprule
|
||||
\textbf{Algorithmus} & \textbf{\textalpha-Helix [\si{\percent}]} & \textbf{\textbeta-Strand [\si{\percent}]} & \textbf{\textbeta-Turn [\si{\percent}]} & \textbf{Coil [\si{\percent}]} \\
|
||||
\midrule
|
||||
K2D3 & 56 & 18 & & \\
|
||||
SELCON3 & 56 & 5 & 17 & 22 \\
|
||||
CONTINLL & 68 & 1 & 14 & 17 \\
|
||||
CDSSTR & 51 & 20 & 15 & 15 \\
|
||||
\midrule
|
||||
SOPMA & 47 & 17 & 9 & 27 \\
|
||||
\midrule
|
||||
PPDK (\textit{Zea mays}) & 47 & 16 & & \\
|
||||
\bottomrule
|
||||
\end{tabular}
|
||||
\end{table}
|
||||
|
||||
\section{Aktivitätsassay}
|
||||
\label{sec:ergebnisse_aktivitaetsassay}
|
||||
Um die Aktivität der gereinigten PPDK zu quantifizieren, wurden zunächst die beiden Fraktionen mit der höchsten \acs{PPDK}-Konzentration -- Fraktion 2 und
|
||||
Fraktion 3 (\SIlist{200;250}{\milli\Molar} Imidazol) -- untersucht (vgl. \ref{sec:aktivitaetsassay}). Hierbei zeigte sich im Vergleich zur Negativkontrolle,
|
||||
dass in beiden Fraktionen eine Aktivität zu verzeichnen ist (siehe Abbildung \ref{abb:aktivitaet}). Da hinsichtlich zukünftiger Kristallisationsexperimente
|
||||
in erster Linie die monodispers vorliegende Fraktion 3 von Interesse ist, wurde von dieser Fraktion eine vollständige Reaktionskinetik aufgenommen. Hierbei ergaben
|
||||
sich durch Anpassen einer Sättigungsfunktion durch nichtlineare Regression an die Michaelis"=Menten"=Gleichung \eqref{eq:mm} für \acs{ATP} und Pyruvat ein \ce{K_M} von
|
||||
\SI[seperr]{50(9)}{\micro\Molar} bzw. \SI[seperr]{270(44)}{\micro\Molar}, sowie ein \ce{V_{max}} von \SI[seperr]{0,095(5)}{\milli\Molar\per\minute} (ATP)
|
||||
\SI[seperr]{0,093(5)}{\milli\Molar\per\minute} (Pyruvat). Auffällig ist die bei höheren \acs{ATP}-Konzentrationen wieder abfallende Reaktionsgeschwindigkeit (vgl. Abbildung \ref{abb:kinetik}).
|
||||
%
|
||||
\begin{align}
|
||||
\label{eq:mm}
|
||||
v &= \frac{V_\text{max} \cdot [S]}{K_m + [S]}
|
||||
\end{align}
|
||||
|
||||
Die Wechselzahl \ce{k_{cat}} wurde anhand von Gleichung \eqref{eq:kcat} aus \ce{V_{max}} und der PPDK-Konzentration von \SI[seperr]{0,38(2)}{\milli\Molar} mit
|
||||
\SI[seperr]{1,63(9)}{\per\second} bestimmt. Die spezifische Aktivität der \acs{PPDK} beträgt \SI[seperr]{0,99(9)}{\Unit\per\milli\gram} und konnte ebenfalls unter Berücksichtigung der
|
||||
Proteinkonzentration mittels Gleichung \ref{eq:spez_akt} berechnet werden.
|
||||
%
|
||||
\begin{align}
|
||||
\label{eq:kcat}
|
||||
k_\text{cat} &= \frac{V_\text{max}}{[E_0]}
|
||||
\end{align}
|
||||
|
||||
Die katalytische Effizient {\ce{k_{cat}}/\ce{K_M}} beträgt \SI[seperr]{32,60(936)}{\milli\Molar\second} (ATP) bzw.
|
||||
\SI[seperr]{6,04(154)}{\milli\Molar\second} (Pyruvat). Eine Auflistung der kinetischen Parameter ist in Tabelle \ref{tab:kinetische_parameter} zu finden.
|
||||
%
|
||||
\begin{align}
|
||||
\label{eq:spez_akt}
|
||||
\text{spez. Aktivität \si{\Unit\per\milli\gram}} &= \frac{V_\text{max}~[\si{\micro\Molar\per\minute}] \cdot V(\text{Ansatz})~[\si{\liter}]}{m(\text{Protein})~[\si{\milli\gram}]}
|
||||
\end{align}
|
||||
|
||||
\begin{table}[htb]
|
||||
\centering
|
||||
\caption[Kinetische Parameter der \acs{PPDK}]{Kinetische Parameter der \acs{PPDK}.}
|
||||
\label{tab:kinetische_parameter}
|
||||
\begin{tabular}{lS[seperr, table-format = 3.3(2)]s}
|
||||
\toprule
|
||||
\textbf{Parameter} & \textbf{Wert} & \textbf{Einheit}\\
|
||||
\midrule
|
||||
\ce{V_{max}} & 0,093(5) & \milli\Molar\per\minute \\
|
||||
\ce{k_{cat}} & 1,63(9) & \per\second \\
|
||||
\midrule
|
||||
\ce{K_m} (ATP) & 50(9) & \micro\Molar \\
|
||||
\ce{K_m} (Pyruvat) & 270(44) & \micro\Molar \\
|
||||
\midrule
|
||||
\ce{k_{cat}}/\ce{K_M} (ATP) & 32,60(936)& \milli\Molar\second \\
|
||||
\ce{k_{cat}}/\ce{K_M} (Pyruvat) & 6,04(154) & \milli\Molar\second \\
|
||||
\midrule
|
||||
spez. Aktivität & 0,99(9) & \Unit\per\milli\gram \\
|
||||
\bottomrule
|
||||
\end{tabular}
|
||||
\end{table}
|
||||
|
||||
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[width=0.7\textwidth,keepaspectratio=true]{./img/ergebnisse/kinetik/aktivitaet.eps}
|
||||
% aktivitaet.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=6 4 466 375
|
||||
\caption[Aktivität der gereinigten PPDK]{Aktivität der gereinigten PPDK. Die Absorption bei \SI{340}{\nano\meter} wurde über einen Zeitraum von \SI{80}{\second}
|
||||
aufgenommen und auf die jeweilige PPDK-Konzentration, sowie das absolute Absorptionsmaximum normiert. Die Kontrolle enthielt anstelle der PPDK ein identisches
|
||||
Volumen Puffer. Die beiden PPDK-Fraktionen weisen eine deutliche Absorptionsabnahme auf, während die Absorption der Kontrolle nahezu unverändert bleibt.}
|
||||
\label{abb:aktivitaet}
|
||||
\end{figure}
|
||||
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\begin{subfigure}[b]{0.45\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/kinetik/atp.eps}
|
||||
% atp.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=2 4 467 375
|
||||
\caption{}
|
||||
\label{abb:kinetik_atp}
|
||||
\end{subfigure}
|
||||
%
|
||||
~
|
||||
%
|
||||
\begin{subfigure}[b]{0.45\textwidth}
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/kinetik/pyruvat.eps}
|
||||
% pyruvat.eps: 0x0 pixel, 300dpi, 0.00x0.00 cm, bb=2 3 467 375
|
||||
\caption{}
|
||||
\label{abb:kinetik_pyruvat}
|
||||
\end{subfigure}
|
||||
\caption[Reaktionskinetik der \acs{PPDK}]{Reaktionskinetik der \acs{PPDK} mit \acs{ATP} (a) und Pyruvat (b) als Substrat. Der inhibierende Effekt von \acs{ATP} auf
|
||||
die im gekoppelten Enzymassay eingesetzte \acs{MDH} ist in (a) ersichtlich. Die Ordinatenachse ist der besseren Übersichtlichkeit wegen logarithmisch skaliert.}
|
||||
\label{abb:kinetik}
|
||||
\end{figure}
|
||||
|
||||
\clearpage
|
||||
|
||||
\section{Homologiemodelle}
|
||||
\label{sec:ergebnisse_homologemodell}
|
||||
Die erzeugten Homologiemodelle (vgl. \ref{sec:erstellung_homologiemodell}) der \acs{PPDK} aus \acs{F. trinervia} zeigen eine gute Übereinstimmung mit
|
||||
den zu Grunde liegenden Templat-Strukturen: \SI{0,708}{\angstrom} für das auf der Struktur aus \acs{C. symbiosum} und \SI{0,705}{\angstrom} für das auf der Struktur aus
|
||||
\textit{Zea mays} basierende Modell. Die Sequenzidentität zur zwischen \acs{F. trinervia} und \textit{Zea mays} beträgt \SI{79}{\percent}, zwischen \acs{F. trinervia} und
|
||||
\acs{C. symbiosum} \SI{55}{\percent}. Das jeweils beste Modell wurde anhand des zDOPE Wertes, sowie der Ergebnisse der PROCHECK-Analyse ausgewählt. Im Ramachandran-Plot
|
||||
zeigt keines der beiden ausgewählten Modelle eine Kombination von ψ- und φ-Winkel, die nicht in einem der erlaubten Bereiche liegt (vgl. Abbildungen \ref{abb:ramachandran_Zm}
|
||||
und \ref{abb:ramachandran_Cs}).
|
||||
|
||||
Anhand der Modelle wurde eine Visualisierung des \textit{Domain-Swiveling} Mechanismus für die \acs{PPDK} aus \acs{F. trinervia} (Abbildung \ref{abb:overlay_homologiemodell}), sowie
|
||||
der \acs{PEP}-Bindestelle (Abbildung \ref{abb:substratbindetasche}) erstellt. Es zeigt sich eine deutliche Torsion der zentralen Phospo-Histidindomäne und des katalytischen
|
||||
His458.
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[width=\textwidth,keepaspectratio=true]{./img/ergebnisse/homologiemodell/swiveling_domain2.png}
|
||||
% swiveling_domain2.png: 2480x976 pixel, 119dpi, 53.13x20.91 cm, bb=0 0 1506 593
|
||||
\caption[Homologiemodelle der \acs{PPDK}]{Homologiemodelle der \acs{PPDK} aus \acs{F. trinervia}. (A) zeigt die \acs{PEP} gebundene Konformation basierend auf der Struktur aus
|
||||
\textit{Zea mays}, (B) das auf \acs{C. symbiosum} basierende Modell. Beide Modelle stellen die jeweiligen Extremkonformationen des \textit{Swiveling-domain} Mechanismus dar. Das
|
||||
an der Übertragung der Phosphatgruppe beteiligte His458 ist als Stabmodell rot eingefärbt und durch einen schwarzen Pfeil markiert. Die einzelnen Domänen der \acs{PPDK} sind ebenfalls
|
||||
farblich hervorgehoben: Nukleotidbindedomäne (grün), Pyruvat/PEP-Bindedomäne (blau), Phosphohistidin-Domäne (gelb), Linker-Peptide (magenta und rot).}
|
||||
\label{abb:overlay_homologiemodell}
|
||||
\end{figure}
|
||||
|
||||
In der Darstellung der modellierten Substratbindestelle sind die für die Koordination von \acs{PEP} wichtigen Reste Arg562, Arg619, Arg669, Glu748, Asp772 und
|
||||
Asn771 sichtbar. Ebenfalls erkennbar ist das als Protonen-Donor fungierende Cys834. Die genannten Reste befinden sich in hoch konservierten Bereichen (siehe Abbildung \ref{abb:ma}).
|
||||
%
|
||||
\begin{figure}[htb]
|
||||
\centering
|
||||
\includegraphics[width=0.8\textwidth,keepaspectratio=true]{./img/ergebnisse/homologiemodell/substratbindung.png}
|
||||
% substratbindung.png: 2480x1681 pixel, 90dpi, 70.00x47.45 cm, bb=0 0 1984 1345
|
||||
\caption[Substratbindetasche für \acs{PEP}]{Substratbindetasche für \acs{PEP}. Die Ansicht zeigt die Bindestelle für \acs{PEP} im
|
||||
Homologiemodell der \acs{PPDK} aus \acs{F. trinervia}. Seitenketten in räumlicher Nähe zum Substrat sind als Linienmodell dargestellt und benannt. Die
|
||||
gestrichelten Linien zeigen mögliche polare Wechselwirkungen mit einer Länge von maximal \SI{3,5}{\angstrom} an.}
|
||||
\label{abb:substratbindetasche}
|
||||
\end{figure}
|
||||
|
||||
|
|
|
|||
|
|
@ -168,10 +168,10 @@
|
|||
|
||||
\subsection{Plasmide}
|
||||
\label{sec:plasmide}
|
||||
Das tatsächlich eingesetzte Plasmid trägt die Bezeichnung pETEV-16b und ist ein Derivat des pET-16b-Vektors,
|
||||
das zusätzlich zwischen der für den Hexa-Histidin-Tag kodierenden Sequenz und dem Zielgen eine für die Schnittstelle
|
||||
der \acs{TEV}-Protease kodierende Sequenz beinhaltet.
|
||||
%
|
||||
% Das tatsächlich eingesetzte Plasmid trägt die Bezeichnung pETEV-16b und ist ein Derivat des pET-16b-Vektors,
|
||||
% das zusätzlich zwischen der für den Hexa-Histidin-Tag kodierenden Sequenz und dem Zielgen eine für die Schnittstelle
|
||||
% der \acs{TEV}-Protease kodierende Sequenz beinhaltet.
|
||||
%
|
||||
\begin{samepage}
|
||||
\begin{tabularx}{\textwidth}{p{0.25\textwidth}p{0.3\textwidth}X}
|
||||
\toprule
|
||||
|
|
@ -423,7 +423,8 @@ vervielfacht werden.
|
|||
\label{sec:inserts_slic}
|
||||
\begin{samepage}
|
||||
Die Synthese von Inserts für eine \ac{SLIC} (vgl.~\ref{sec:slic}) wurde mittels einer \ac{PCR} nach untenstehendem
|
||||
Ansatz durchgeführt. Als Primer kamen die in \ref{sec:synthetische_oligonukleotide} aufgeführten Oligonukleotide zum Einsatz.
|
||||
Ansatz durchgeführt. Als Primer kamen die in \ref{sec:synthetische_oligonukleotide} aufgeführten Oligonukleotide zum Einsatz. Diese
|
||||
wurden so gewählt, dass jeweils \SI{30}{\bp} zum Zielvektor und \SI{20}{\bp} zum Zielgen homolog sind.
|
||||
\begin{description}
|
||||
\item[\ac{PCR}-Ansatz] \hfill \\
|
||||
\SI{1}{\micro\liter} DNA (Templat) \\
|
||||
|
|
@ -518,7 +519,7 @@ Zur Generierung der 5'-Überhänge wurden \SI{1000}{\nano\gram} Vektor- bzw. Ins
|
|||
ad \SI{35}{\micro\liter} \acs{ddH2O}
|
||||
\end{description}
|
||||
%
|
||||
Ziel war ein Dau von etwa \SI{30}{\bp} Länge. Aus der Exonukleaseaktivität der T4-\acs{DNA}-Polymerase von
|
||||
Ziel war ein Dau von etwa \SI{30}{\bp} Länge. Aus der Exonukleaseaktivität der T4"=\acs{DNA}"=Polymerase von
|
||||
\SI{10}{\bp\per\minute} bei \SI{22}{\celsius} ergab sich eine Inkubationsdauer von \SI{40}{\minute}. Die Reaktion wurde
|
||||
durch Zugabe von $\frac{1}{10}$ des Ansatzvolumens \SI{10}{\milli\Molar} \ac{dCTP} gestoppt.
|
||||
|
||||
|
|
@ -611,7 +612,8 @@ einem Proteingemisch isolieren. Im Rahmen dieser Arbeit wurden HisTrap$^\text{\t
|
|||
eingesetzt. Als Säulenmaterial dient quervernetzte Agarose an welche \acf{IDA} immobilisiert ist. \ce{Ni2+}-Ionen werden von \acs{IDA} dreifach koordinativ
|
||||
komplexiert, so dass drei Koordinationsstellen für die Interaktion Poly-Histidin-markierter Proteine mit den immobilisierten \ce{Ni2+}-Ionen zur
|
||||
Verfügung stehen. Die Elution der spezifisch an das Säulenmaterial gebundenen Proteine erfolgt über die Zugabe von Imidazol, welches als Strukturanalogon zu
|
||||
Histidin in der Lage ist, dieses kompetitiv aus der Bindung mit \ce{Ni2+} zu verdrängen.
|
||||
Histidin in der Lage ist, dieses kompetitiv aus der Bindung mit \ce{Ni2+} zu verdrängen. Das nachfolgend beschriebene Reinigungsprotokoll wurde nach \citet{Nakanishi2003,Chastain1996}
|
||||
adaptiert.
|
||||
|
||||
Die wie in \ref{sec:zellaufschluss} beschrieben aufgeschlossenen Zellen wurden zunächst bei \SI{10000}{\xg} und \SI{15}{\celsius} für \SI{30}{\minute}
|
||||
zentrifugiert um Zelltrümmer und \textit{Inclusion-Bodies} zu entfernen. Der Überstand wurde anschließend für \SI{1}{\hour} bei \SI{100000}{\xg} und \SI{15}{\celsius}
|
||||
|
|
@ -716,16 +718,25 @@ Pipettierschema für drei kleine Gele (\SI{9}{\centi\meter}~\texttimes~\SI{10}{\
|
|||
\begin{table}[htb]
|
||||
\centering
|
||||
\caption{Pipettierschema für SDS-Gele}
|
||||
\begin{tabular}{lll}
|
||||
\begin{tabular}{lSsSs}
|
||||
\toprule
|
||||
& {\textbf{Trenngel}} & {\textbf{Sammelgel}}\\
|
||||
& \multicolumn{2}{c}{\textbf{Trenngel}} & \multicolumn{2}{c}{\textbf{Sammelgel}}\\
|
||||
\midrule
|
||||
\acs{AA/BAA} 30 & \SI{10,9}{\milli\liter} & \SI{2}{\milli\liter}\\
|
||||
Trenn-/Sammelgelpuffer & \SI{13,2}{\milli\liter} & \SI{2,5}{\milli\liter}\\
|
||||
\acs{ddH2O} & \SI{8,6}{\milli\liter} & \SI{7,4}{\milli\liter}\\
|
||||
\acs{TEMED} & \SI{16,5}{\micro\liter} & \SI{12,3}{\micro\liter}\\
|
||||
\acs{APS} \SI{10}{\percent} (w/v) & \SI{112,2}{\micro\liter} & \SI{129}{\micro\liter}\\
|
||||
\acs{AA/BAA} 30 & 10,9 & \milli\liter & 2 & \milli\liter \\
|
||||
Trenn-/Sammelgelpuffer & 13,2 & \milli\liter & 2,5 & \milli\liter \\
|
||||
\acs{ddH2O} & 8,6 & \milli\liter & 7,4 & \milli\liter \\
|
||||
\acs{TEMED} & 16,5 & \micro\liter & 12,3 & \micro\liter \\
|
||||
\acs{APS} \SI{10}{\percent} (w/v) & 112,2 & \micro\liter & 129 & \micro\liter \\
|
||||
\bottomrule
|
||||
% \toprule
|
||||
% & {\textbf{Trenngel}} & & {\textbf{Sammelgel}} & \\
|
||||
% \midrule
|
||||
% \acs{AA/BAA} 30 & 10,9 & \milli\liter & 2 & \milli\liter}\\
|
||||
% Trenn-/Sammelgelpuffer & 13,2 & \milli\liter & 2,5 & \milli\liter}\\
|
||||
% \acs{ddH2O} & 8,6 & \milli\liter & 7,4 & \milli\liter}\\
|
||||
% \acs{TEMED} & 16,5 & \micro\liter & 12,3 & \micro\liter}\\
|
||||
% \acs{APS} \SI{10}{\percent} (w/v) & 112,2 & \micro\liter & 129 & \micro\liter}\\
|
||||
% \bottomrule
|
||||
\end{tabular}
|
||||
\label{tab:pipettierschema_sds_page}
|
||||
\end{table}
|
||||
|
|
@ -794,13 +805,13 @@ Die \ac{CD}-Spektroskopie kann für die Analyse von Proteinsekundärstrukturen e
|
|||
\SIrange{160}{250}{\nano\meter} aufgenommen. Man macht sich dabei zu Nutze, dass in diesem Bereich die n~\textrightarrow~\textpi$^*$ bzw.
|
||||
\textpi~\textrightarrow~\textpi$^*$-Übergänge der Peptidbindung liegen. Durch ihre Chiralität reagiert das \ac{CD}-Spektrum eines Peptids sehr
|
||||
empfindlich auf Änderungen der Sekundärstruktur \citep{Greenfield2007}. Die Messungen erfolgten in CD-Puffer bei einer Proteinkonzentration
|
||||
von \SI{0,2}{\milli\gram\per\milli\liter}. Von den Rohdaten wurde das Pufferspektrum abgezogen und unter Berücksichtigung von Konzentration und
|
||||
von \SI{0,1}{\milli\gram\per\milli\liter}. Von den Rohdaten wurde das Pufferspektrum abgezogen und unter Berücksichtigung von Konzentration und
|
||||
Molekulargewicht in molaren \ac{CD} (\textDelta\textepsilon) umgerechnet.
|
||||
|
||||
\subsection{Aktivitätsassay}
|
||||
\label{sec:aktivitaetsassay}
|
||||
Um die Aktivität der gereinigten \acs{PPDK} zu bestimmten, wurde ein Aktivitätsassay \citep{Salahas1990} durchgeführt. Hierbei wurde \ac{PEP}-Bildung
|
||||
in einem gekoppelten Enzymassay über den \acs{NADH}-Verbrauch verfolgt. Der geschwindigkeitsbestimmende Schritt ist in diesem Fall die Umsetzung
|
||||
in einem gekoppelten Enzymassay über den \acs{NADH}"=Verbrauch verfolgt. Der geschwindigkeitsbestimmende Schritt ist in diesem Fall die Umsetzung
|
||||
von Pyruvat, anorganischem Phosphat und \acs{ATP} zu \acl{PEP}, \acs{AMP} und Pyrophosphat \eqref{eq:aktivitaet_1}. Nachfolgend wird \acl{PEP} durch
|
||||
die \acl{PEPCase} über eine \textbeta-Carboxylierung mit Hydrogencarbonat zu Oxalacetat umgesetzt \eqref{eq:aktivitaet_2}. Letzteres wird durch eine
|
||||
NAD-abhängige Malatdehydrogenase zu Malat reduziert \eqref{eq:aktivitaet_3}.
|
||||
|
|
@ -816,7 +827,7 @@ NAD-abhängige Malatdehydrogenase zu Malat reduziert \eqref{eq:aktivitaet_3}.
|
|||
}
|
||||
\end{align}
|
||||
%
|
||||
Der \acs{NADH}-Verbrauch kann photometrisch über die Abnahme der \acs{NADH}-spezifischen Extinktion bei \SI{340}{\nano\meter} verfolgt werden. Da der
|
||||
Der \acs{NADH}-Verbrauch kann photometrisch über die Abnahme der \acs{NADH}"=spezifischen Extinktion bei \SI{340}{\nano\meter} verfolgt werden. Da der
|
||||
Verbrauch eines \acs{NADH}-Moleküls gleichbedeutend ist mit der Bildung eines \acl{PEP}-Moleküls durch die \acs{PPDK}, kann aus dem Verbrauch an
|
||||
\acs{NADH} direkt auf die Bildungsrate von \acl{PEP} und somit auf die Aktivität der \acs{PPDK} geschlossen werden.
|
||||
|
||||
|
|
@ -842,8 +853,9 @@ $\epsilon_{340} = \SI{6,3e3}{\per\Molar\per\centi\meter}$ \citep{McComb1976} und
|
|||
|
||||
\subsection{Analyse von \ac{CD}-Spektren}
|
||||
\label{sec:analyse_cd_spektren}
|
||||
Die Analyse der aufgenommenen CD-Spektrem (siehe \ref{sec:cd_spektroskopie}) erfolgte mit dem Programmpaket \textit{CDPro} \citep{Sreerama2000}, sowie
|
||||
dem Programm \textit{K2D3} \citep{Louis-Jeune2011}. Die Analyse erfolgte jeweils in einem Wellenlängenbereich von \SIrange{190}{240}{\nano\meter}.
|
||||
Aufgenommene CD-Spektrem (siehe \ref{sec:cd_spektroskopie}) wurden mit dem Programmpaket \textit{CDPro} \citep{Sreerama2000}, sowie
|
||||
dem Programm \textit{K2D3} \citep{Louis-Jeune2011} untersucht. Die Analyse erfolgte jeweils in einem Wellenlängenbereich von \SIrange{190}{240}{\nano\meter}.
|
||||
Für das Programmpaket \textit{CDPro} wurde die Datenbank SMP56 mit 43 löslichen und 13 Membranproteinen als Grundlage der Vorhersage verwendet.
|
||||
Die berechneten Sekundärstrukturanteile wurden dann mit mit einer \textit{ab initio} Vorhersage, welche mit \acs{SOPMA} \citep{Geourjon1995a}
|
||||
berechnet wurde verglichen. Zusätzlich wurden die Sekundärstrukturanteile aus der bereits bekannten \acs{PPDK}-Struktur aus \textit{Zea mays}
|
||||
herangezogen.
|
||||
|
|
@ -859,8 +871,9 @@ Zur Visualisierung der wahrscheinlichen dreidimensionalen Struktur der \acs{PPDK
|
|||
wurden Homologiemodelle erstellt. Diese basieren auf den aus \textit{Zea mays} (PDB: 1VBG/1VBH) und \textit{Chlostridium symbiosum} (PDB: 1KBL) bekannten
|
||||
Extremkonformationen des putativen \textit{Domain-swiveling}-Mechanismus.
|
||||
|
||||
Die Homologiemodelle wurden mit dem Programm \textit{Modeller} \citep{Sali1993a} aus den jeweiligen Templaten und der Primärstruktur der
|
||||
Jeweils fünf Homologiemodelle wurden mit dem Programm \textit{Modeller} \citep{Sali1993a} aus den jeweiligen Templaten und der Primärstruktur der
|
||||
\acs{PPDK} aus \acs{F. trinervia} \citep{Rosche1990} generiert. Im Falle des von \textit{Zea mays} abgeleiteten Modells wurde als Vorlage die Pyruvat gebundene Struktur
|
||||
verwendet (PDB: 1VBH). Lücken in der dreidimensionalen Struktur wurden anhand der ungebundenen Struktur (PDB: 1VBG) ergänzt.
|
||||
verwendet (PDB: 1VBH). Lücken in der dreidimensionalen Struktur wurden anhand der ungebundenen Struktur (PDB: 1VBG) ergänzt. Um die Substratbindetasche möglichst
|
||||
exakt zu nachzubilden, wurde das in der Kristallstruktur 1VBH vorhandene \acs{PEP}-Molekül, sowie das \ce{Mg2+}-Ion beim Modellieren berücksichtigt.
|
||||
|
||||
Die Evaluation der erstellten Modelle erfolgte mit dem Programmpaket \textit{PROCHECK} \citep{Laskowski1993,Laskowski1996}.
|
||||
|
|
|
|||
|
|
@ -1,2 +1 @@
|
|||
\addchap{Zusammenfassung}
|
||||
<Zusammenfassung>
|
||||
\addchap{Zusammenfassung}
|
||||
|
|
|
|||
4171
library.bib
4171
library.bib
File diff suppressed because it is too large
Load diff
|
|
@ -4,7 +4,7 @@ img_extIsRegExp=false
|
|||
img_extensions=.eps .jpg .jpeg .png .pdf .ps .fig .gif
|
||||
kileprversion=2
|
||||
kileversion=2.1.2
|
||||
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|
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|
||||
masterDocument=
|
||||
name=Masterarbeit
|
||||
pkg_extIsRegExp=false
|
||||
|
|
@ -126,6 +126,76 @@ mode=
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|||
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|
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|
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|
||||
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|
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|
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|
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BIN
masterarbeit.pdf
Normal file
BIN
masterarbeit.pdf
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Binary file not shown.
|
|
@ -12,7 +12,8 @@
|
|||
oneside, % Einsitiges Dokument
|
||||
headinclude, % Kopf dem Satzspiegel zurechnen wg. lebendem Kolumnentitel
|
||||
ngerman, % Benutzte Sprachen werden global definiert (benötigt von
|
||||
english % siunitx)
|
||||
english, % siunitx)
|
||||
abstracton
|
||||
]{scrreprt} % Dokumentenklasse
|
||||
|
||||
\usepackage{scrhack}
|
||||
|
|
@ -30,6 +31,8 @@
|
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%\directlua{fonts.protrusions.setups.default.factor=.5}
|
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\defaultfontfeatures{Ligatures=TeX} %,Numbers=OldStyle}% ,Scale=MatchLowercase}
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\usepackage{libertineotf}
|
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\setmonofont[Scale=1]{Inconsolata}
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%\setmonofont[Scale=0.9]{LinMonoO}
|
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|
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|
|
@ -41,15 +44,19 @@
|
|||
\setkomafont{pagehead}{\normalfont\sffamily} % Serifenlose Schrift im Seitenkopf
|
||||
\setheadsepline{.4pt} % Breite der Linie unter dem Seitenkopf
|
||||
\chead{} % Unterdrücken der Kolumnentitel in der Mitte des Seitenkopfes
|
||||
%\cfoot{} % Unterdrücken der Seitenzahl unten auf der Seite
|
||||
%\cfoot{} % Unterdrücken der Seitenzahl unten auf der Seite
|
||||
\ihead{\headmark} % Seitenkopf innen: Kolumnentitel
|
||||
%\ohead{\pagemark} % Seitenkopf außen: Seitenzahl
|
||||
%\ohead{\pagemark} % Seitenkopf außen: Seitenzahl
|
||||
|
||||
\pdfcompresslevel=9
|
||||
\usepackage{setspace} % Paket für das Setzen des Zeilenabstandes
|
||||
\onehalfspacing % 1,5facher Zeilenabstand
|
||||
\usepackage{acronym} % Abkürzungen und Abkürzungsverzeichnis
|
||||
\usepackage[alsoload=synchem,locale=DE,obeyall,math-micro=\textmu,text-micro=\textmu]{siunitx}
|
||||
\usepackage{setspace} % Paket für das Setzen des Zeilenabstandes
|
||||
\onehalfspacing % 1,5facher Zeilenabstand
|
||||
\usepackage[printonlyused]{acronym} % Abkürzungen und Abkürzungsverzeichnis
|
||||
\usepackage[alsoload=synchem,
|
||||
locale=DE,
|
||||
obeyall,
|
||||
math-micro=\textmu,
|
||||
text-micro=\textmu]{siunitx}
|
||||
|
||||
\newunit{\bp}{bp} % Definition eigener Einheiten: Basenpaare
|
||||
\newunit{\kb}{kb} % Definition eigener Einheiten: Kilo-Basenpaare
|
||||
|
|
@ -67,13 +74,17 @@
|
|||
\usepackage{tabularx} % Tabellen mit fester Breite
|
||||
\usepackage{longtable} % Seitenüberspannende Tabellen
|
||||
\usepackage{booktabs} % 'Hübsche' Tabellen ohne vertikale Linien
|
||||
\usepackage{threeparttable} % Fußnoten in Tabellen
|
||||
\usepackage{multirow} % Mehrzeilige Tabellen
|
||||
\usepackage{array}
|
||||
\usepackage{graphicx} % Grafikerweiterung
|
||||
\usepackage{float} % Exakte Positionierung von Grafiken
|
||||
\usepackage{tikz} % Grafikbibliothek, u.a. benötigt von mhchem + pgf
|
||||
\usepackage{gnuplot-lua-tikz}
|
||||
\usepackage[version=3]{mhchem} % Darstellung chemischer Summenformeln
|
||||
\mhchemoptions{arrows=pgf} % Fix für die Darstellung von Pfeilen mit unicode-math
|
||||
\usepackage{texshade}
|
||||
\usepackage{url}
|
||||
\usepackage{texshade} % Sequenzalignments
|
||||
\usepackage{url} % Automatische Formatierung von URLs
|
||||
\usepackage{natbib} % Naturwissenschaftliche Zitierungsstile
|
||||
\usepackage[format=plain,footnotesize]{caption}[2008/08/24]
|
||||
\usepackage{subcaption} % Untergeordnete Abbildungen
|
||||
|
|
@ -81,6 +92,9 @@
|
|||
\usepackage[fixlanguage]{babelbib} % Sprachanpassung für Literaturverzeichnis
|
||||
\selectbiblanguage{ngerman} % s.o.
|
||||
|
||||
\usepackage{etoolbox}
|
||||
\patchcmd{\bibsetup}{\interlinepenalty=5000}{\interlinepenalty=10000}{}{}
|
||||
|
||||
\usepackage{hyperref} % PDF-Eigenschaften (s.u.)
|
||||
\hypersetup{
|
||||
pdfencoding=auto,
|
||||
|
|
@ -192,7 +206,8 @@
|
|||
|
||||
\nocite{Li2007}
|
||||
|
||||
\bibliographystyle{natdin}
|
||||
|
||||
\bibliographystyle{natdin_edit}
|
||||
\bibliography{./library}
|
||||
|
||||
\include{./inc/anhang}
|
||||
|
|
|
|||
|
|
@ -1,5 +1,5 @@
|
|||
>F. trinervia
|
||||
TAKKRVFTFGKGRSEGNRDMKSLLGGKGANLAEMSSIGLSVPPGLTISTE
|
||||
KKRVFTFGKGRSEGNRDMKSLLGGKGANLAEMSSIGLSVPPGLTISTE
|
||||
ACEEYQQNGKSLPPGLWDEISEGLDYVQKEMSASLGDPSKPLLLSVRSGA
|
||||
AISMPGMMDTVLNLGLNDEVVAGLAGKSG-ARFAYDSYRRFLDMFGNVVM
|
||||
GIPHSLFDEKLEQMKAEKGIHLDTDLTAADLKDLVEKYKNVYVEA-KGEK
|
||||
|
|
@ -18,7 +18,7 @@ TMIEIPRAALIAEEIGKEADFFSFGTNDLTQMTFGYSRDDVGKFLQIYLA
|
|||
QGILQHDPFEVIDQKGVGQLIKMATEKGRAANPSLKVGICGEHGGEPSSV
|
||||
AFFDGVGLDYVSCSPFRVPIARLAAAQVI---V
|
||||
>F. brownii
|
||||
TTKKRVFTFGKGNSEGNKDMKSLLGGKGANLAEMASIGLSVPPGLTISTE
|
||||
KKRVFTFGKGNSEGNKDMKSLLGGKGANLAEMASIGLSVPPGLTISTE
|
||||
ACEEYQQNGKKLPPGLWDEILEGLQYVQKEMSASLGDPSKALLLSVRSGA
|
||||
AISMPGMMDTVLNLGLNDEVVDGLAAKSG-ARFAYDSYRRFLDMFGNVVM
|
||||
GIPHSLFDEKLEQMKAEKGIHLDTDLTAADLKDLAEQYKNVYVEA-KGEK
|
||||
|
|
@ -37,7 +37,7 @@ TMIEIPRAALIADEIAKEAEFFSFGTNDLTQMTFGYSRDDVGKFLPIYLS
|
|||
QGILQHDPFEVLDQKGVGQLIKMATEKGRAANPNLKVGICGEHGGEPSSV
|
||||
AFFDGVGLDYVSCSPFRVPIARLAAAQVV---V
|
||||
>Zea mays
|
||||
TTKKRVFHFGKGKSEGNKTMKELLGGKGANLAEMASIGLSVPPGFTVSTE
|
||||
KKRVFHFGKGKSEGNKTMKELLGGKGANLAEMASIGLSVPPGFTVSTE
|
||||
ACQQYQDAGCALPAGLWAEIVDGLQWVEEYMGATLGDPQRPLLLSVRSGA
|
||||
AVSMPGMMDTVLNLGLNDEVAAGLAAKSG-ERFAYDSFRRFLDMFGNVVM
|
||||
DIPRSLFEEKLEHMKESKGLKNDTDLTASDLKELVGQYKEVYLSA-KGEP
|
||||
|
|
@ -56,7 +56,7 @@ TMIEIPRAALVADEIAEQAEFFSFGTNDLTQMTFGYSRDDVGKFIPVHLA
|
|||
QGILQHDPFEVLDQRGVGELVKFATERGRKARPNLKVGICGEHGGEPSSV
|
||||
AFFAKAGLDFVSCSPFRVPIARLAAAQVL---V
|
||||
>C. symbiosum
|
||||
MA-KWVYKF----EEGNASMRNLLGGKGCNLAEMTILGMPIPQGFTVTTE
|
||||
-KWVYKF----EEGNASMRNLLGGKGCNLAEMTILGMPIPQGFTVTTE
|
||||
ACTEYYNSGKQITQEIQDQIFEAITWLEELNGKKFGDTEDPLLVSVRSGA
|
||||
RASMPGMMDTILNLGLNDVAVEGFAKKTGNPRFAYDSYRRFIQMYSDVVM
|
||||
EVPKSHFEKIIDAMKEEKGVHFDTDLTADDLKELAEKFKAVYKEAMNGEE
|
||||
|
|
|
|||
76
misc/ma_as_aln.fa~
Normal file
76
misc/ma_as_aln.fa~
Normal file
|
|
@ -0,0 +1,76 @@
|
|||
>F. trinervia
|
||||
TAKKRVFTFGKGRSEGNRDMKSLLGGKGANLAEMSSIGLSVPPGLTISTE
|
||||
ACEEYQQNGKSLPPGLWDEISEGLDYVQKEMSASLGDPSKPLLLSVRSGA
|
||||
AISMPGMMDTVLNLGLNDEVVAGLAGKSG-ARFAYDSYRRFLDMFGNVVM
|
||||
GIPHSLFDEKLEQMKAEKGIHLDTDLTAADLKDLVEKYKNVYVEA-KGEK
|
||||
FPTDPKKQLELAVNAVFDSWDSPRANKYRSINQITGLKGTAVNIQSMVFG
|
||||
NMGNTSGTGVLFTRNPSTGEKKLYGEFLINAQGEDVVAGIRTPEDLGTME
|
||||
TCMPEAYKELVENCEILERHYKDMMDIEFTVQENRLWMLQCRTGKRTGKG
|
||||
AVRIAVDMVNEGLIDTRTAIKRVETQHLDQLLHPQFEDPSAYK-SHVVAT
|
||||
GLPASPGAAVGQVCFSAEDAETWHAQGKSAILVRTETSPEDVGGMHAAAG
|
||||
ILTARGGMTSHAAVVARGWGKCCVSGCADIRVNDDMKIFTIGDRVIKEGD
|
||||
WLSLNGTTGEVILGKQLLAPPAMSNDLEIFMSWADQARRLKVMANADTPN
|
||||
DALTARNNGAQGIGLCRTEHMFFASDERIKAVRKMIMAVTPEQRKVALDL
|
||||
LLPYQRSDFEGIFRAMDGLPVTIRLLDPPLHEFLPEGDLEHIVNELAVDT
|
||||
GMSADEIYSKIENLSEVNPMLGFRGCRLGISYPELTEMQVRAIFQAAVSM
|
||||
TN-QGVTVIPEIMVPLVGTPQELRHQISVIRGVAANVFAEMGVTLEYKVG
|
||||
TMIEIPRAALIAEEIGKEADFFSFGTNDLTQMTFGYSRDDVGKFLQIYLA
|
||||
QGILQHDPFEVIDQKGVGQLIKMATEKGRAANPSLKVGICGEHGGEPSSV
|
||||
AFFDGVGLDYVSCSPFRVPIARLAAAQVI---V
|
||||
>F. brownii
|
||||
TTKKRVFTFGKGNSEGNKDMKSLLGGKGANLAEMASIGLSVPPGLTISTE
|
||||
ACEEYQQNGKKLPPGLWDEILEGLQYVQKEMSASLGDPSKALLLSVRSGA
|
||||
AISMPGMMDTVLNLGLNDEVVDGLAAKSG-ARFAYDSYRRFLDMFGNVVM
|
||||
GIPHSLFDEKLEQMKAEKGIHLDTDLTAADLKDLAEQYKNVYVEA-KGEK
|
||||
FPTDPKKQLELAVNAVFDSWDSPRANKYRSINQITGLKGTAVNIQCMVFG
|
||||
NMGNTSGTGVLFTRNPSTGEKKLYGEFLVNAQGEDVVAGIRTPEDLVTME
|
||||
TCMPEAYRELVENCVILERHYKDMMDIEFTVQENRLWMLQCRTGKRTGKG
|
||||
AVRIAVDMVNEGLIDTRTAIKRVETQHLDQLLHPQFENPSAYK-SHVVAT
|
||||
GLPASPGAAVGQVVFSAEDAETWHAQGKSAILVRTETSPEDVGGMHAAAG
|
||||
ILTARGGMTSHAAVVARGWGKCCVSGCADIRVNDDMKVFTIGDRVIKEGD
|
||||
WLSLNGSTGEVILGKQLLAPPAMSNDLETFMSWADQARRLKVMANADTPN
|
||||
DALTARNNGAQGIGLCRTEHMFFASDERIKAVRKMIMAVTPEQRKAALDL
|
||||
LLPYQRSDFEGIFRAMDGLPVTIRLLDPPLHEFLPEGDLEHIVNELTADT
|
||||
GMSKDEIYSRIEKLSEVNPMLGFRGCRLGISYPELTEMQVRAIFQAAVSM
|
||||
NN-QGVTVIPEIMVPLVGTPQELRHQIGVIRGVAANVFAEMGLTLEYKVG
|
||||
TMIEIPRAALIADEIAKEAEFFSFGTNDLTQMTFGYSRDDVGKFLPIYLS
|
||||
QGILQHDPFEVLDQKGVGQLIKMATEKGRAANPNLKVGICGEHGGEPSSV
|
||||
AFFDGVGLDYVSCSPFRVPIARLAAAQVV---V
|
||||
>Zea mays
|
||||
TTKKRVFHFGKGKSEGNKTMKELLGGKGANLAEMASIGLSVPPGFTVSTE
|
||||
ACQQYQDAGCALPAGLWAEIVDGLQWVEEYMGATLGDPQRPLLLSVRSGA
|
||||
AVSMPGMMDTVLNLGLNDEVAAGLAAKSG-ERFAYDSFRRFLDMFGNVVM
|
||||
DIPRSLFEEKLEHMKESKGLKNDTDLTASDLKELVGQYKEVYLSA-KGEP
|
||||
FPSDPKKQLELAVLAVFNSWESPRAKKYRSINQITGLRGTAVNVQCMVFG
|
||||
NMGNTSGTGVLFTRNPNTGEKKLYGEFLVNAQGEDVVAGIRTPEDLDAMK
|
||||
NLMPQAYDELVENCNILESHYKEMQDIEFTVQENRLWMLQCRTGKRTGKS
|
||||
AVKIAVDMVNEGLVEPRSAIKMVEPGHLDQLLHPQFENPSAYK-DQVIAT
|
||||
GLPASPGAAVGQVVFTAEDAEAWHSQGKAAILVRAETSPEDVGGMHAAVG
|
||||
ILTERGGMTSHAAVVARWWGKCCVSGCSGIRVNDAEKLVTIGSHVLREGE
|
||||
WLSLNGSTGEVILGKQPLSPPALSGDLGTFMAWVDDVRKLKVLANADTPD
|
||||
DALTARNNGAQGIGLCRTEHMFFASDERIKAVRQMIMAPTLELRQQALDR
|
||||
LLTYQRSDFEGIFRAMDGLPVTIRLLDHPSYEFLPEGNIEDIVSELCAET
|
||||
GANQEDALARIEKLSEVNPMLGFRGCRLGISYPELTEMQARAIFEAAIAM
|
||||
TN-QGVQVFPEIMVPLVGTPQELGHQVTLIRQVAEKVFANVGKTIGYKVG
|
||||
TMIEIPRAALVADEIAEQAEFFSFGTNDLTQMTFGYSRDDVGKFIPVHLA
|
||||
QGILQHDPFEVLDQRGVGELVKFATERGRKARPNLKVGICGEHGGEPSSV
|
||||
AFFAKAGLDFVSCSPFRVPIARLAAAQVL---V
|
||||
>C. symbiosum
|
||||
MA-KWVYKF----EEGNASMRNLLGGKGCNLAEMTILGMPIPQGFTVTTE
|
||||
ACTEYYNSGKQITQEIQDQIFEAITWLEELNGKKFGDTEDPLLVSVRSGA
|
||||
RASMPGMMDTILNLGLNDVAVEGFAKKTGNPRFAYDSYRRFIQMYSDVVM
|
||||
EVPKSHFEKIIDAMKEEKGVHFDTDLTADDLKELAEKFKAVYKEAMNGEE
|
||||
FPQEPKDQLMGAVKAVFRSWDNPRAIVYRRMNDIPGDWGTAVNVQTMVFG
|
||||
NKGETSGTGVAFTRNPSTGEKGIYGEYLINAQGEDVVAGVRTPQPITQLE
|
||||
NDMPDCYKQFMDLAMKLEKHFRDMQDMEFTIEEGKLYFLQTRNGKRTAPA
|
||||
ALQIACDLVDEGMITEEEAVVRIEAKSLDQLLHPTF-NPAALKAGEVIGS
|
||||
ALPASPGAAAGKVYFTADEAKAAHEKGERVILVRLETSPEDIEGMHAAEG
|
||||
ILTVRGGMTSHAAVVARGMGTCCVSGCGEIKINEEAKTFELGGHTFAEGD
|
||||
YISLDGSTGKIYKGDIETQEASVSGSFERIMVWADKFRTLKVRTNADTPE
|
||||
DTLNAVKLGAEGIGLCRTEHMFFEAD-RIMKIRKMILSDSVEAREEALNE
|
||||
LIPFQKGDFKAMYKALEGRPMTVRYLDPPLHEFVPHTEEE--QAELAKNM
|
||||
GLTLAEVKAKVDELHEFNPMMGHRGCRLAVTYPEIAKMQTRAVMEAAIEV
|
||||
KEETGIDIVPEIMIPLVGEKKELKFVKDVVVEVAEQVKKEKGSDMQYHIG
|
||||
TMIEIPRAALTADAIAEEAEFFSFGTNDLTQMTFGFSRDDAGKFLDSYYK
|
||||
AKIYESDPFARLDQTGVGQLVEMAVKKGRQTRPGLKCGICGEHGGDPSSV
|
||||
EFCHKVGLNYVSCSPFRVPIARLAAAQAALNNK
|
||||
Loading…
Add table
Reference in a new issue