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| - | ==Crystal structure of a de novo designed parallel four-helix coiled coil, 4-KE-4== | + | ==Crystal structure of a de novo designed parallel four-helix coiled coil, 4-KE-4.== |
| | <StructureSection load='6xy1' size='340' side='right'caption='[[6xy1]], [[Resolution|resolution]] 1.50Å' scene=''> | | <StructureSection load='6xy1' size='340' side='right'caption='[[6xy1]], [[Resolution|resolution]] 1.50Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[6xy1]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6XY1 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6XY1 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6xy1]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6XY1 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6XY1 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=NH2:AMINO+GROUP'>NH2</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.5Å</td></tr> |
| - | <tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACE:ACETYL+GROUP'>ACE</scene></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACE:ACETYL+GROUP'>ACE</scene>, <scene name='pdbligand=NH2:AMINO+GROUP'>NH2</scene></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6xy1 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6xy1 OCA], [http://pdbe.org/6xy1 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6xy1 RCSB], [http://www.ebi.ac.uk/pdbsum/6xy1 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6xy1 ProSAT]</span></td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=6xy1 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6xy1 OCA], [https://pdbe.org/6xy1 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6xy1 RCSB], [https://www.ebi.ac.uk/pdbsum/6xy1 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6xy1 ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
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| | </StructureSection> | | </StructureSection> |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Edgell, C L]] | + | [[Category: Synthetic construct]] |
| - | [[Category: Savery, N J]] | + | [[Category: Edgell CL]] |
| - | [[Category: Woolfson, D N]] | + | [[Category: Savery NJ]] |
| - | [[Category: Coiled coil]] | + | [[Category: Woolfson DN]] |
| - | [[Category: De novo protein]]
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| - | [[Category: Parallel]]
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| - | [[Category: Tetramer]]
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| Structural highlights
Publication Abstract from PubMed
De novo-designed protein domains are increasingly being applied in biotechnology, cell biology, and synthetic biology. Therefore, it is imperative that these proteins be robust to superficial changes; i.e., small changes to their amino acid sequences should not cause gross structural changes. In turn, this allows properties such as stability and solubility to be tuned without affecting structural attributes like tertiary fold and quaternary interactions. Reliably designed proteins with predictable behaviors may then be used as scaffolds to incorporate function, e.g., through the introduction of features for small-molecule, metal, or macromolecular binding, and enzyme-like active sites. Generally, achieving this requires the starting protein fold to be well understood. Herein, we focus on designing alpha-helical coiled coils, which are well studied, widespread, and often direct protein-protein interactions in natural systems. Our initial investigations reveal that a previously designed parallel, homotetrameric coiled coil, CC-Tet, is not robust to sequence changes that were anticipated to maintain its structure. Instead, the alterations switch the oligomeric state from tetramer to trimer. To improve the robustness of designed homotetramers, additional sequences based on CC-Tet were produced and characterized in solution and by X-ray crystallography. Of these updated sequences, one is robust to truncation and to changes in surface electrostatics; we call this CC-Tet*. Variants of the general CC-Tet* design provide a set of homotetrameric coiled coils with unfolding temperatures in the range from 40 to >95 degrees C. We anticipate that these will be of use in applications requiring robust and well-defined tetramerization domains.
Robust De Novo-Designed Homotetrameric Coiled Coils.,Edgell CL, Savery NJ, Woolfson DN Biochemistry. 2020 Mar 17;59(10):1087-1092. doi: 10.1021/acs.biochem.0c00082., Epub 2020 Mar 9. PMID:32133841[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Edgell CL, Savery NJ, Woolfson DN. Robust De Novo-Designed Homotetrameric Coiled Coils. Biochemistry. 2020 Mar 17;59(10):1087-1092. doi: 10.1021/acs.biochem.0c00082., Epub 2020 Mar 9. PMID:32133841 doi:http://dx.doi.org/10.1021/acs.biochem.0c00082
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