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| | ==Crystal Structure of Engineered Trimeric Cortexillin-1 Coiled-Coil Variant== | | ==Crystal Structure of Engineered Trimeric Cortexillin-1 Coiled-Coil Variant== |
| - | <StructureSection load='4j4a' size='340' side='right' caption='[[4j4a]], [[Resolution|resolution]] 1.65Å' scene=''> | + | <StructureSection load='4j4a' size='340' side='right'caption='[[4j4a]], [[Resolution|resolution]] 1.65Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4j4a]] is a 12 chain structure with sequence from [http://en.wikipedia.org/wiki/Atcc_11735 Atcc 11735]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4J4A OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4J4A FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4j4a]] is a 12 chain structure with sequence from [https://en.wikipedia.org/wiki/Dictyostelium_discoideum Dictyostelium discoideum]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4J4A OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4J4A FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</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]] 1.6499Å</td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">ctxA, DDB_G0289483 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=44689 ATCC 11735])</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=PO4:PHOSPHATE+ION'>PO4</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=4j4a FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4j4a OCA], [http://pdbe.org/4j4a PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4j4a RCSB], [http://www.ebi.ac.uk/pdbsum/4j4a PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4j4a 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=4j4a FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4j4a OCA], [https://pdbe.org/4j4a PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4j4a RCSB], [https://www.ebi.ac.uk/pdbsum/4j4a PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4j4a ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[http://www.uniprot.org/uniprot/CTXA_DICDI CTXA_DICDI]] Acting-bundling protein. When linked to F-actin the actin filaments form preferentially anti-parallel bundles that associate into meshworks. Plays a major role in cytokinesis. Negatively regulates cortical localization of rapgap1.<ref>PMID:18039932</ref> | + | [https://www.uniprot.org/uniprot/CTXA_DICDI CTXA_DICDI] Acting-bundling protein. When linked to F-actin the actin filaments form preferentially anti-parallel bundles that associate into meshworks. Plays a major role in cytokinesis. Negatively regulates cortical localization of rapgap1.<ref>PMID:18039932</ref> |
| - | <div style="background-color:#fffaf0;">
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| - | == Publication Abstract from PubMed ==
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| - | Coiled coils are well suited to drive subunit oligomerization and are widely used in applications ranging from basic research to medicine. The optimization of these applications requires a detailed understanding of the molecular determinants that control of coiled-coil formation. Although many of these determinants have been identified and characterized in great detail, a puzzling observation is that their presence does not necessarily correlate with the oligomerization state of a given coiled-coil structure. Thus, other determinants must play a key role. To address this issue, we recently investigated the unrelated coiled-coil domains from GCN4, ATF1 and cortexillin-1 as model systems. We found that well-known trimer-specific oligomerization-state determinants, such as the distribution of isoleucine residues at heptad-repeat core positions or the trimerization motif Arg-h-x-x-h-Glu (where h = hydrophobic amino acid; x = any amino acid), switch the peptide's topology from a dimer to a trimer only when inserted into the trigger sequence, a site indispensable for coiled-coil formation. Because high-resolution structural information could not be obtained for the full-length, three-stranded cortexillin-1 coiled coil, we here report the detailed biophysical and structural characterization of a shorter variant spanning the trigger sequence using circular dichroism, anatytical ultracentrifugation and x-ray crystallography. We show that the peptide forms a stable alpha-helical trimer in solution. We further determined the crystal structure of an optimised variant at a resolution of 1.65 A, revealing that the peptide folds into a parallel, three-stranded coiled coil. The two complemented R-IxxIE trimerization motifs and the additional hydrophobic core isoleucine residue adopt the conformations seen in other extensively characterized parallel, three-stranded coiled coils. These findings not only confirm the structural basis for the switch in oligomerization state from a dimer to a trimer observed for the full-length cortexillin-1 coiled-coil domain, but also provide further evidence for a general link between oligomerization-state specificity and trigger-sequence function.
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| - | Structural basis for the oligomerization-state switch from a dimer to a trimer of an engineered cortexillin-1 coiled-coil variant.,Bjelic S, Wieser M, Frey D, Stirnimann CU, Chance MR, Jaussi R, Steinmetz MO, Kammerer RA PLoS One. 2013 May 14;8(5):e63370. doi: 10.1371/journal.pone.0063370. Print 2013. PMID:23691037<ref>PMID:23691037</ref>
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| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| - | </div>
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| - | <div class="pdbe-citations 4j4a" style="background-color:#fffaf0;"></div>
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| | == References == | | == References == |
| | <references/> | | <references/> |
| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Atcc 11735]] | + | [[Category: Dictyostelium discoideum]] |
| - | [[Category: Bjelic, S]] | + | [[Category: Large Structures]] |
| - | [[Category: Kammerer, R A]] | + | [[Category: Bjelic S]] |
| - | [[Category: Steinmetz, M O]] | + | [[Category: Kammerer RA]] |
| - | [[Category: Coiled-coil]] | + | [[Category: Steinmetz MO]] |
| - | [[Category: De novo protein]]
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| - | [[Category: Trimer]]
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