Team:KAIST-Korea/Project/Modeling

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__NOTOC__
__NOTOC__
{{:Team:KAIST/header}}
{{:Team:KAIST/header}}
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== Single chain antibody structural alignment ==
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<table width="100%" border="0" cellpadding="20px">
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=== Protocol ===
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<tr>
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There are four steps to compare structure of single chain antibody and FGF binding domain of FGFR. First step is taking variable region sequences of antibodies. Next step is combining these variable region sequences with linker sequence to make single chain antibody sequence. Third step is predicting the structure of single chain antibody with structure prediction program like modeler. Final step is to structural align these structures of antibodies with structure of FGF binding domain of FGFR(PDB ID : 1EVT).
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<td valign="top" width="75%">
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==== Data source ====
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<span style=font-size:30px><font face="Maiandra GD"><b> Modeling </b></font></span>
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Single chain antibody is the combination of variable regions of known antibodies with linker sequence which can bind to the antigens. We need to know the V<sub>L</sub> and V<sub>H</sub> sequences to make single chain antibody. The source of these antibody sequences are NCBI, Uniprot and RCSB PDB. NCBI and Uniprot provide the single chain sequence of variable regions (V<sub>L</sub> and V<sub>H</sub>) and antigen binding fragments (F<sub>ab</sub>). RCSB provide the structure of the complexes of antigenbinding fragment which binds to its antigens. But we only need the sequence of variable region. So we get the last 120~150 reside and assume them as the variable region. And data from RCSB contain not only sequence of antibody, but also antigens. S we filter them based on label of files.to get heavy chains and light chains of antibody.
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<br>
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----
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==== Single chain antibody synthesis ====
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<br>
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We combine the antibody variable region sequences with order of V<sub>H</sub>-linker-V<sub>L</sub> to make single chain antibody sequence. The sequence of linker is GGGGSGGGGS
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&nbsp;&nbsp;Leaving cell transmit signal through chemical modifications of signal-related molecules like phosphorylation, methylation, acetylation or ubiquitination. And these chemical modifications are basically chemical reactions. Therefore, we can assume signal transduction pathway as sequential chemical reactions. We applied this assumption to signal transduction initiated by modified FGFR-antigen binding. Signal transduction follows these steps.
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<br>
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==== Structure Prediction ====
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<br>
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==== Structure Alignment ====
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# Antigen binds to the single chain antibody of modified FGFR and form FGFR-complex(FGFRC).
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=== Result===
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# Two FGFR-complexes bind each other and form FGFR-complex-dimer(FGFRCD).
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{| class="sortable wikitable" border="0" align="center" style="border: 1px solid #999; background-color:#FFFFFF"
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# FGFRCD phosphorylate each other and form FGFR-complex-dimer-phosphorylate (FGFRCDnP)
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|-align="left" bgcolor="#CCCCCC"
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# FGFRCDnP phosphorylate STAT1 to for STAT1P.
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! Name
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# Two STAT1Ps bind each other and form STAT1P-dimer.
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!Core Residues||Core RMSD||Raw Score||P-value
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# STAT1P-dimer near cell surface diffuse to inside of nucleus of yeast.
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|-
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# STAT1P-dimer in nucleus binds to GAS element and activate GFP gene.
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| 2VXT
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<br>
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| 95
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<br>
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| 2.676
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&nbsp;&nbsp;And these steps may be formulated as these reactions.
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| 84.306
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<br>
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| 0.000222
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<html>
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|-
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<img src = "https://static.igem.org/mediawiki/2010/b/bd/Modeling1.png" width = 400>
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| 2VXU
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</html>
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| 98
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<br>
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| 2.563
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<br>
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| 92.118
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&nbsp;&nbsp;And we can expect that the concentration of FGFRCDnP is constant at equilibrium state. The concentration of FGFRCDnP is<br>
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| 0.0000823
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<html>
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|-
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<img src = "https://static.igem.org/mediawiki/2010/a/a6/KAISTModeling2.png" width = 200>
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| 2VXV
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</html>
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| 77
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<br>
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| 2.64
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<br>
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| 52.269
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&nbsp;&nbsp;And the concentration of FGFRCD is
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| 0.0644
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<br>
-
|-
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<html>
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| 2ZKH
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<img src = "https://static.igem.org/mediawiki/2010/5/5e/KAISTModeling3.png" width = 150>
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| 96
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</html>
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| 2.855
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<br>
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| 93.411
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<br>
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| 0.001
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&nbsp;&nbsp;And the concentration of FGFRC is
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|-
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<br>
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| 3AAZ
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<html>
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| 84
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<img src = "https://static.igem.org/mediawiki/2010/c/c0/KAISTModeling4.png" width = 200>
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| 4.197
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</html>
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| 68.078
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<br>
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| 0.0642
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<br>
-
|-
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&nbsp;&nbsp;Therefore, the concentration of FGFRCDnP as the function of concentration of FGFR [FGFR], that of antigen [antigen] and phosphorylation number n is<br>
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| 3D69
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<html>
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| 137
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<img src = "https://static.igem.org/mediawiki/2010/0/02/KAISTModeling5.png" width = 500>
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| 12.224
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</html>
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| 98.134
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<br>
-
| 0.00541
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<br>
-
|-
+
-
| 3EO9
+
-
| 80
+
-
| 2.282
+
-
| 84.665
+
-
| 0.00295
+
-
|-
+
-
| 3EOA
+
-
| 116
+
-
| 10.402
+
-
| 98.544
+
-
| 0.0547
+
-
|-
+
-
| 3EOB
+
-
| 68
+
-
| 4.685
+
-
| 56.357
+
-
| 0.3753
+
-
|-
+
-
| 3EYV
+
-
| 85
+
-
| 4.704
+
-
| 66.669
+
-
| 0.1653
+
-
|-
+
-
| 3FMG
+
-
| 81
+
-
| 2.629
+
-
| 73.298
+
-
| 0.0129
+
-
|-
+
-
| 3FOG
+
-
| 90
+
-
| 7.243
+
-
| 74.031
+
-
| 0.0785
+
-
|-
+
-
| 3G6D
+
-
| 72
+
-
| 4.403
+
-
| 69.774
+
-
| 0.1281
+
-
|-
+
-
| 3GBM
+
-
| 119
+
-
| 10.331
+
-
| 115.929
+
-
| 0.0314
+
-
|-
+
-
| 3GBN
+
-
| 77
+
-
| 2.788
+
-
| 57.204
+
-
| 0.0592
+
-
|-
+
-
| 3GHB
+
-
| 101
+
-
| 12.861
+
-
| 81.189
+
-
| 0.3984
+
-
|-
+
-
| 3GHE
+
-
| 96
+
-
| 3.302
+
-
| 109.208
+
-
| 0.00117
+
-
|-
+
-
| 3GI8
+
-
| 127
+
-
| 6.837
+
-
| 113.467
+
-
| 0.00038
+
-
|-
+
-
| 3GI9
+
-
| 52
+
-
| 3.534
+
-
| 43.56
+
-
| 0.166
+
-
|-
+
-
| 3GIZ
+
-
| 89
+
-
| 8.217
+
-
| 68.702
+
-
| 0.3172
+
-
|-
+
-
| 3GK8
+
-
| 91
+
-
| 6.84
+
-
| 84.048
+
-
| 0.00811
+
-
|-
+
-
| 3GKW
+
-
| 74
+
-
| 3.205
+
-
| 47.096
+
-
| 0.1215
+
-
|-
+
-
| 3GNM
+
-
| 135
+
-
| 4.739
+
-
| 103.812
+
-
| 0.00000297
+
-
|-
+
-
| 3GO1
+
-
| 103
+
-
| 8.842
+
-
| 77.631
+
-
| 0.2587
+
-
|-
+
-
| 3GRW
+
-
| 107
+
-
| 9.819
+
-
| 108.405
+
-
| 0.0449
+
-
|-
+
-
| 3H42
+
-
| 69
+
-
| 4.389
+
-
| 59.165
+
-
| 0.3421
+
-
|-
+
-
| 3HC0
+
-
| 67
+
-
| 3.065
+
-
| 40.982
+
-
| 0.1028
+
-
|-
+
-
| 3HC3
+
-
| 67
+
-
| 3.173
+
-
| 49.109
+
-
| 0.0944
+
-
|-
+
-
| 3HC4
+
-
| 36
+
-
| 1.998
+
-
| 42.538
+
-
| 0.206
+
-
|-
+
-
| 3HI5
+
-
| 77
+
-
| 3.222
+
-
| 58.096
+
-
| 0.1832
+
-
|-
+
-
| 3HI6
+
-
| 81
+
-
| 3.646
+
-
| 63.526
+
-
| 0.0394
+
-
|-
+
-
| 3HMW
+
-
| 83
+
-
| 4.159
+
-
| 62.598
+
-
| 0.1976
+
-
|-
+
-
| 3HMX
+
-
| 70
+
-
| 5.215
+
-
| 62.814
+
-
| 0.2228
+
-
|-
+
-
| 3HNT
+
-
| 84
+
-
| 2.62
+
-
| 74.666
+
-
| 0.00118
+
-
|-
+
-
| 3HNV
+
-
| 84
+
-
| 2.998
+
-
| 77.17
+
-
| 0.00374
+
-
|-
+
-
| 3HR5
+
-
| 78
+
-
| 4.158
+
-
| 58.026
+
-
| 0.2978
+
-
|-
+
-
| 3I50
+
-
| 65
+
-
| 2.206
+
-
| 52.252
+
-
| 0.1156
+
-
|-
+
-
| 3I9G
+
-
| 84
+
-
| 5.026
+
-
| 64.155
+
-
| 0.1506
+
-
|-
+
-
| 3IU3
+
-
| 75
+
-
| 3.192
+
-
| 56.006
+
-
| 0.2552
+
-
|-
+
-
| 3IXT
+
-
| 78
+
-
| 2.951
+
-
| 68.627
+
-
| 0.0378
+
-
|-
+
-
| 3KDM
+
-
| 87
+
-
| 4.273
+
-
| 65.722
+
-
| 0.1683
+
-
|-
+
-
| 3KS0
+
-
| 81
+
-
| 3.039
+
-
| 55.562
+
-
| 0.0192
+
-
|-
+
-
| 3KYK
+
-
| 88
+
-
| 4.728
+
-
| 68.681
+
-
| 0.1016
+
-
|-
+
-
| 3KYM
+
-
| 85
+
-
| 6.52
+
-
| 66.102
+
-
| 0.5135
+
-
|-
+
-
| 3L1O
+
-
| 93
+
-
| 2.545
+
-
| 85.909
+
-
| 0.0014
+
-
|-
+
-
| 3L5W
+
-
| 89
+
-
| 4.925
+
-
| 72.178
+
-
| 0.0637
+
-
|-
+
-
| 3L5X
+
-
| 88
+
-
| 4.796
+
-
| 65.262
+
-
| 0.1111
+
-
|-
+
-
| 3L5Y
+
-
| 64
+
-
| 4.689
+
-
| 65.227
+
-
| 0.2007
+
-
|-
+
-
| 3L95
+
-
| 73
+
-
| 5.105
+
-
| 70.312
+
-
| 0.0515
+
-
|-
+
-
| 3LMJ
+
-
| 76
+
-
| 2.699
+
-
| 56.627
+
-
| 0.0237
+
-
|-
+
-
| 3LQA
+
-
| 75
+
-
| 8.371
+
-
| 58.374
+
-
| 0.8315
+
-
|-
+
-
| 3LS4
+
-
| 79
+
-
| 2.458
+
-
| 72.68
+
-
| 0.00694
+
-
|-
+
-
| 3LS5
+
-
| 81
+
-
| 2.443
+
-
| 73.043
+
-
| 0.0014
+
-
|-
+
-
| 3LZF
+
-
| 96
+
-
| 2.468
+
-
| 93.292
+
-
| 0.00155
+
-
|-
+
-
| 3MLR
+
-
| 71
+
-
| 2.574
+
-
| 64.979
+
-
| 0.0165
+
-
|-
+
-
| 3MLS
+
-
| 75
+
-
| 2.534
+
-
| 57.386
+
-
| 0.0202
+
-
|-
+
-
| 3MLU
+
-
| 127
+
-
| 10.025
+
-
| 73.766
+
-
| 0.0305
+
-
|-
+
-
| 3MLV
+
-
| 75
+
-
| 2.454
+
-
| 62.321
+
-
| 0.0218
+
-
|-
+
-
| 3MLW
+
-
| 83
+
-
| 8.94
+
-
| 59.029
+
-
| 0.4846
+
-
|-
+
-
| 3MLX
+
-
| 118
+
-
| 11.485
+
-
| 105.391
+
-
| 0.0961
+
-
|-
+
-
| 3MLY
+
-
| 84
+
-
| 4.704
+
-
| 63.962
+
-
| 0.1023
+
-
|-
+
-
| 3MLZ
+
-
| 82
+
-
| 3.995
+
-
| 64.563
+
-
| 0.123
+
-
|-
+
-
| 3MUG
+
-
| 79
+
-
| 2.827
+
-
| 68.72
+
-
| 0.0223
+
-
|-
+
-
| 3MXV
+
-
| 125
+
-
| 10.857
+
-
| 121.911
+
-
| 0.01
+
-
|-
+
-
| 3MXW
+
-
| 102
+
-
| 11.262
+
-
| 78.348
+
-
| 0.4213
+
-
|--bgcolor="#C3C4D8"
+
-
| 16A1
+
-
| 86
+
-
| 2.139
+
-
| 89.893
+
-
| 0.00443
+
-
|-
+
-
|}
+
-
 
+
-
===Result Analysis===
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Latest revision as of 05:22, 16 August 2010

 

Modeling


  Leaving cell transmit signal through chemical modifications of signal-related molecules like phosphorylation, methylation, acetylation or ubiquitination. And these chemical modifications are basically chemical reactions. Therefore, we can assume signal transduction pathway as sequential chemical reactions. We applied this assumption to signal transduction initiated by modified FGFR-antigen binding. Signal transduction follows these steps.

  1. Antigen binds to the single chain antibody of modified FGFR and form FGFR-complex(FGFRC).
  2. Two FGFR-complexes bind each other and form FGFR-complex-dimer(FGFRCD).
  3. FGFRCD phosphorylate each other and form FGFR-complex-dimer-phosphorylate (FGFRCDnP)
  4. FGFRCDnP phosphorylate STAT1 to for STAT1P.
  5. Two STAT1Ps bind each other and form STAT1P-dimer.
  6. STAT1P-dimer near cell surface diffuse to inside of nucleus of yeast.
  7. STAT1P-dimer in nucleus binds to GAS element and activate GFP gene.



  And these steps may be formulated as these reactions.


  And we can expect that the concentration of FGFRCDnP is constant at equilibrium state. The concentration of FGFRCDnP is


  And the concentration of FGFRCD is


  And the concentration of FGFRC is


  Therefore, the concentration of FGFRCDnP as the function of concentration of FGFR [FGFR], that of antigen [antigen] and phosphorylation number n is
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