Team:Grenoble-EMSE-LSU/Documentation/Biobricks

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<h3>Structure</h3>
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<h3>Structure</h3><br><br>
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<p>Below is a comparison of the 3D structures of monomeric dsRed and KillerRed:<br><br>
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                        <p>In order to understand why KillerRed has its unique properties it is necessary to look at its structure. The protein is remarkably similar to other fluorescent proteins like GFP <em>(Aequorea victoria></em> and dsRed <em>(Discosoma striata)</em>, featuring a beta-barrel housing a central alpha helix with the fluorescent chromophore at its center. Normally the chromophore is protected from the outside environment by the protein shell, but this isn't the case with KillerRed.
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<p align="center"><img src="https://static.igem.org/mediawiki/2013/8/8c/DsRed_alongside_KillerRed.png" alt="dsRed and KillerRed protein structures."></p><br>
<p align="center"><img src="https://static.igem.org/mediawiki/2013/8/8c/DsRed_alongside_KillerRed.png" alt="dsRed and KillerRed protein structures."></p><br>
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                         <em><strong>Source: RCSB protein database entries <a href="http://www.rcsb.org/pdb/explore/explore.do?structureId=2WIQ">2WIQ</a> and <a href="http://www.rcsb.org/pdb/explore/explore.do?structureId=2VAD">2VAD</a>.</strong><em></p><br>
                         <em><strong>Source: RCSB protein database entries <a href="http://www.rcsb.org/pdb/explore/explore.do?structureId=2WIQ">2WIQ</a> and <a href="http://www.rcsb.org/pdb/explore/explore.do?structureId=2VAD">2VAD</a>.</strong><em></p><br>
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KillerRed is a 240 amino acid protein with a 3D structure similar to other fluorescent proteins, with an eleven-strand beta-barrel surrounding an alpha-helix containing the chromophore, source of the protein's optical properties.<br>
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<p>KillerRed is a 240 amino acid protein with a 3D structure similar to other fluorescent proteins, with an eleven-strand beta-barrel surrounding an alpha-helix containing the chromophore, source of the protein's optical properties.<br>
KillerRed has a DsRed-type chromophore formed with residues 67Q (glutamine), 68Y (tyrosine), and 69G (glycine), to make QYG. The corresponding coding sequence can be found at the code segment CAGTACGGC.<br><br>
KillerRed has a DsRed-type chromophore formed with residues 67Q (glutamine), 68Y (tyrosine), and 69G (glycine), to make QYG. The corresponding coding sequence can be found at the code segment CAGTACGGC.<br><br>
The interesting properties of the protein are directly related to a unique structural difference among fluorescent proteins, consisting in an open channel linking the chromophore to the environment outside the protein. According to litterature, this is the reason KillerRed is able to produce 1000-fold more reactive oxygen species compared to EGFP which is another ROS-producing fluorescent protein.
The interesting properties of the protein are directly related to a unique structural difference among fluorescent proteins, consisting in an open channel linking the chromophore to the environment outside the protein. According to litterature, this is the reason KillerRed is able to produce 1000-fold more reactive oxygen species compared to EGFP which is another ROS-producing fluorescent protein.
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<li><h2>References</h2>
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                                <p>[1] Sergei Pletnev et al., Structural Basis for Phototoxicity of the Genetically Encoded Photosensitizer KillerRed, <em>THE JOURNAL OF BIOLOGICAL CHEMISTRY</em> vol. 284, no. 46, pp. 32028–32039, November 13, 2009.<br>
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                                  [2] M.E. Bulina et al., A genetically encoded photosensitizer, <em>Nature Biotechnology</em>, January 2006.<br>
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                                  [3] J.J. Tabor et al., Multichromatic Control of Gene Expression in <em>Escherichia coli</em>, <em>Journal of Molecular Biology</em>, 2011.<br>
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                                  [4] K.E. McGinness et al., Engineering Controllable Protein Degradation, <em>Molecular Cell</em>, June 2006.
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Revision as of 15:43, 19 September 2013

Grenoble-EMSE-LSU, iGEM


Grenoble-EMSE-LSU, iGEM