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| | ==C-terminal domain of CTP1L endolysin mutant V195P that reduces autoproteolysis== | | ==C-terminal domain of CTP1L endolysin mutant V195P that reduces autoproteolysis== |
| - | <StructureSection load='4cu2' size='340' side='right' caption='[[4cu2]], [[Resolution|resolution]] 2.11Å' scene=''> | + | <StructureSection load='4cu2' size='340' side='right'caption='[[4cu2]], [[Resolution|resolution]] 2.11Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[4cu2]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4CU2 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4CU2 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[4cu2]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Clostridium_phage_phiCTP1 Clostridium phage phiCTP1]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4CU2 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=4CU2 FirstGlance]. <br> |
| - | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[4cu5|4cu5]]</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]] 2.11Å</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=4cu2 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4cu2 OCA], [http://pdbe.org/4cu2 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=4cu2 RCSB], [http://www.ebi.ac.uk/pdbsum/4cu2 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=4cu2 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=4cu2 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4cu2 OCA], [https://pdbe.org/4cu2 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=4cu2 RCSB], [https://www.ebi.ac.uk/pdbsum/4cu2 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=4cu2 ProSAT]</span></td></tr> |
| | </table> | | </table> |
| - | <div style="background-color:#fffaf0;">
| + | == Function == |
| - | == Publication Abstract from PubMed == | + | [https://www.uniprot.org/uniprot/D9ZNF3_9CAUD D9ZNF3_9CAUD] |
| - | The bacteriophage PhiCD27 is capable of lysing Clostridium difficile, a pathogenic bacterium that is a major cause for nosocomial infection. A recombinant CD27L endolysin lyses C. difficile in vitro, and represents a promising alternative as a bactericide. To better understand the lysis mechanism, we have determined the crystal structure of an autoproteolytic fragment of the CD27L endolysin. The structure covers the C-terminal domain of the endolysin, and represents a novel fold that is identified in a number of lysins that target Clostridia bacteria. The structure indicates endolysin cleavage occurs at the stem of the linker connecting the catalytic domain with the C-terminal domain. We also solved the crystal structure of the C-terminal domain of a slow cleaving mutant of the CTP1L endolysin that targets C. tyrobutyricum. Two distinct dimerization modes are observed in the crystal structures for both endolysins, despite a sequence identity of only 22% between the domains. The dimers are validated to be present for the full length protein in solution by right angle light scattering, small angle X-ray scattering and cross-linking experiments using the cross-linking amino acid p-benzoyl-L-phenylalanine (pBpa). Mutagenesis on residues contributing to the dimer interfaces indicates that there is a link between the dimerization modes and the autocleavage mechanism. We show that for the CTP1L endolysin, there is a reduction in lysis efficiency that is proportional to the cleavage efficiency. We propose a model for endolysin triggering, where the extended dimer presents the inactive state, and a switch to the side-by-side dimer triggers the cleavage of the C-terminal domain. This leads to the release of the catalytic portion of the endolysin, enabling the efficient digestion of the bacterial cell wall.
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| - | The CD27L and CTP1L Endolysins Targeting Clostridia Contain a Built-in Trigger and Release Factor.,Dunne M, Mertens HD, Garefalaki V, Jeffries CM, Thompson A, Lemke EA, Svergun DI, Mayer MJ, Narbad A, Meijers R PLoS Pathog. 2014 Jul 24;10(7):e1004228. doi: 10.1371/journal.ppat.1004228., eCollection 2014 Jul. PMID:25058163<ref>PMID:25058163</ref>
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| - | | + | |
| - | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| - | </div>
| + | |
| - | <div class="pdbe-citations 4cu2" style="background-color:#fffaf0;"></div>
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| - | == References ==
| + | |
| - | <references/>
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Dunne, M]] | + | [[Category: Clostridium phage phiCTP1]] |
| - | [[Category: Garefalaki, V]] | + | [[Category: Large Structures]] |
| - | [[Category: Jeffries, C M]] | + | [[Category: Dunne M]] |
| - | [[Category: Lemke, E A]] | + | [[Category: Garefalaki V]] |
| - | [[Category: Mayer, M J]] | + | [[Category: Jeffries CM]] |
| - | [[Category: Meijers, R]] | + | [[Category: Lemke EA]] |
| - | [[Category: Mertens, H D.T]] | + | [[Category: Mayer MJ]] |
| - | [[Category: Narbad, A]] | + | [[Category: Meijers R]] |
| - | [[Category: Svergun, D I]] | + | [[Category: Mertens HDT]] |
| - | [[Category: Thompson, A]] | + | [[Category: Narbad A]] |
| - | [[Category: Autoproteolysis]] | + | [[Category: Svergun DI]] |
| - | [[Category: Bacterial lysis]] | + | [[Category: Thompson A]] |
| - | [[Category: Hydrolase]]
| + | |
