Difference between revisions of "Team:Shenzhen SFLS/Protein Engineering"

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<p><blockquote>  P185<sup>her2/neu</sup>, member of the HER receptor family, is chosen as the antigen to be mutated in our project. We used several programs to help us have a better overall understanding of the antigen. Since only the extracellular domain can be recognized and bound by an antibody, we would only mutate and express the extracellular domain (ECD). Also, some results below only cover ECD and its mutants.</blockquote></p>
 
<p><blockquote>  P185<sup>her2/neu</sup>, member of the HER receptor family, is chosen as the antigen to be mutated in our project. We used several programs to help us have a better overall understanding of the antigen. Since only the extracellular domain can be recognized and bound by an antibody, we would only mutate and express the extracellular domain (ECD). Also, some results below only cover ECD and its mutants.</blockquote></p>
 
<center><a href="https://static.igem.org/mediawiki/2015/9/90/PRdePE_M0_result.txt">Numerical_result_of_M0</a></center>
 
<center><a href="https://static.igem.org/mediawiki/2015/9/90/PRdePE_M0_result.txt">Numerical_result_of_M0</a></center>
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</br><i><center>Fig 3.1 .Polarity scale of p185her2/neu ECD, predicted by ProtScale, Zimmerman polarity</center></i>
 
</br><i><center>Fig 3.1 .Polarity scale of p185her2/neu ECD, predicted by ProtScale, Zimmerman polarity</center></i>
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</br><i><center>Fig 3.2 .Hydrophobicity scale of p185her2/neu ECD, predicted by ProtScale, Hphob. / Eisenberg et al.</i></center>
 
</br><i><center>Fig 3.2 .Hydrophobicity scale of p185her2/neu ECD, predicted by ProtScale, Hphob. / Eisenberg et al.</i></center>
 
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</br><i><center><blockquote><blockquote><blockquote><blockquote><blockquote><blockquote>Fig 3.2 .Hydrophobicity scale of p185her2/neu ECD, predicted by ProtScale, Hphob. / Eisenberg et al.Fig 3.3. Secondary structure and solvent accessibility prediction of p185her2/neu ECD, predicted by PredictProtein. The black scale plate above with numbers indicates the number of residues. Corresponding to the residue numbers, the stripes below the scale plate indicate the secondary structure of the region covered. Blue stripes indicate strand region, while red stripes indicate helix region. Results of the first and second row are respectively predicted by RePROF (new and experimental) and PROFsec (veteran). Results of the third row indicate whether the corresponding region is exposed or buried. Blue stripes indicate that the regions covered are exposed, and yellow stripes indicate that the regions covered are buried.</blockquote></blockquote></blockquote></blockquote></blockquote></blockquote></center></i>
 
</br><i><center><blockquote><blockquote><blockquote><blockquote><blockquote><blockquote>Fig 3.2 .Hydrophobicity scale of p185her2/neu ECD, predicted by ProtScale, Hphob. / Eisenberg et al.Fig 3.3. Secondary structure and solvent accessibility prediction of p185her2/neu ECD, predicted by PredictProtein. The black scale plate above with numbers indicates the number of residues. Corresponding to the residue numbers, the stripes below the scale plate indicate the secondary structure of the region covered. Blue stripes indicate strand region, while red stripes indicate helix region. Results of the first and second row are respectively predicted by RePROF (new and experimental) and PROFsec (veteran). Results of the third row indicate whether the corresponding region is exposed or buried. Blue stripes indicate that the regions covered are exposed, and yellow stripes indicate that the regions covered are buried.</blockquote></blockquote></blockquote></blockquote></blockquote></blockquote></center></i>
 
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</br><i><center>Fig 3.5. The scFv-EPI interface. EPI consists of the p185her2/neu ECD subdomain I (residues 1~172) and the first C2 module (residues 173~192) of subdomain II, which are respectively colored in green and yellow. Only the three loops containing the epitope are drawn. The light chain (VL) and heavy chain (VH) are colored in blue and red respectively. Other parts of scFv chA21 are colored in grey. A. The stereo overview of the scFv-EPI interface. B. The three loops containing the epitope of EPI lie on the chA21 surface. The important residues on EPI are shown as sticks and labeled with black characters. C-F, the detailed hydrogen bonding interaction network between chA21 and EPI. The hydrogen bonds are drawn as grey dashes. The residues are labeled, and characters in the parentheses indicate which CDR (chA21 scFv) or loop (EPI) the residues belong to. In panels B-F, the oxygen, nitrogen, and sulfur atoms are colored red , blue and orange, respectively.</br></i></center>
 
</br><i><center>Fig 3.5. The scFv-EPI interface. EPI consists of the p185her2/neu ECD subdomain I (residues 1~172) and the first C2 module (residues 173~192) of subdomain II, which are respectively colored in green and yellow. Only the three loops containing the epitope are drawn. The light chain (VL) and heavy chain (VH) are colored in blue and red respectively. Other parts of scFv chA21 are colored in grey. A. The stereo overview of the scFv-EPI interface. B. The three loops containing the epitope of EPI lie on the chA21 surface. The important residues on EPI are shown as sticks and labeled with black characters. C-F, the detailed hydrogen bonding interaction network between chA21 and EPI. The hydrogen bonds are drawn as grey dashes. The residues are labeled, and characters in the parentheses indicate which CDR (chA21 scFv) or loop (EPI) the residues belong to. In panels B-F, the oxygen, nitrogen, and sulfur atoms are colored red , blue and orange, respectively.</br></i></center>
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Revision as of 13:08, 18 September 2015

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3.1 Protein engineering

P185her2/neu, member of the HER receptor family, is chosen as the antigen to be mutated in our project. We used several programs to help us have a better overall understanding of the antigen. Since only the extracellular domain can be recognized and bound by an antibody, we would only mutate and express the extracellular domain (ECD). Also, some results below only cover ECD and its mutants.

Numerical_result_of_M0

Fig 3.1 .Polarity scale of p185her2/neu ECD, predicted by ProtScale, Zimmerman polarity

Fig 3.2 .Hydrophobicity scale of p185her2/neu ECD, predicted by ProtScale, Hphob. / Eisenberg et al.


Fig 3.2 .Hydrophobicity scale of p185her2/neu ECD, predicted by ProtScale, Hphob. / Eisenberg et al.Fig 3.3. Secondary structure and solvent accessibility prediction of p185her2/neu ECD, predicted by PredictProtein. The black scale plate above with numbers indicates the number of residues. Corresponding to the residue numbers, the stripes below the scale plate indicate the secondary structure of the region covered. Blue stripes indicate strand region, while red stripes indicate helix region. Results of the first and second row are respectively predicted by RePROF (new and experimental) and PROFsec (veteran). Results of the third row indicate whether the corresponding region is exposed or buried. Blue stripes indicate that the regions covered are exposed, and yellow stripes indicate that the regions covered are buried.


Fig 3.5. The scFv-EPI interface. EPI consists of the p185her2/neu ECD subdomain I (residues 1~172) and the first C2 module (residues 173~192) of subdomain II, which are respectively colored in green and yellow. Only the three loops containing the epitope are drawn. The light chain (VL) and heavy chain (VH) are colored in blue and red respectively. Other parts of scFv chA21 are colored in grey. A. The stereo overview of the scFv-EPI interface. B. The three loops containing the epitope of EPI lie on the chA21 surface. The important residues on EPI are shown as sticks and labeled with black characters. C-F, the detailed hydrogen bonding interaction network between chA21 and EPI. The hydrogen bonds are drawn as grey dashes. The residues are labeled, and characters in the parentheses indicate which CDR (chA21 scFv) or loop (EPI) the residues belong to. In panels B-F, the oxygen, nitrogen, and sulfur atoms are colored red , blue and orange, respectively.


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