5w3w: Difference between revisions
New page: '''Unreleased structure''' The entry 5w3w is ON HOLD Authors: Hiblot, J., Gotthard, G., Jacquet, P., Daude, D., Bergonzi, C., Chabriere, E., Elias, M. Description: Crystal structure of... |
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The | ==Crystal structure of SsoPox AsD6 mutant (V27A-Y97W-L228M-W263M) - open form== | ||
<StructureSection load='5w3w' size='340' side='right'caption='[[5w3w]], [[Resolution|resolution]] 2.95Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5w3w]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Saccharolobus_solfataricus Saccharolobus solfataricus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5W3W OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5W3W FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.95Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CO:COBALT+(II)+ION'>CO</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=FE2:FE+(II)+ION'>FE2</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=KCX:LYSINE+NZ-CARBOXYLIC+ACID'>KCX</scene></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=5w3w FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5w3w OCA], [https://pdbe.org/5w3w PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5w3w RCSB], [https://www.ebi.ac.uk/pdbsum/5w3w PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5w3w ProSAT]</span></td></tr> | |||
</table> | |||
== Function == | |||
[https://www.uniprot.org/uniprot/PHP_SACS2 PHP_SACS2] Has a low paraoxonase activity. Also active, but with a lower activity, against other organo-phosphorus insecticides such as Dursban, Coumaphos, pNP-butanoate or parathion.<ref>PMID:15909078</ref> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
The redesign of enzyme active sites to alter their function or specificity is a difficult yet appealing challenge. Here we used a structure-based design approach to engineer the lactonase SsoPox from Sulfolobus solfataricus into a phosphotriesterase. The five best variants were characterized and their structure was solved. The most active variant, alphasD6 (V27A-Y97W-L228M-W263M) demonstrates a large increase in catalytic efficiencies over the wild-type enzyme, with increases of 2,210-fold, 163-fold, 58-fold, 16-fold against methyl-parathion, malathion, ethyl-paraoxon, and methyl-paraoxon, respectively. Interestingly, the best mutants are also capable of degrading fensulfothion, which is reported to be an inhibitor for the wild-type enzyme, as well as others that are not substrates of the starting template or previously reported W263 mutants. The broad specificity of these engineered variants makes them promising candidates for the bioremediation of organophosphorus compounds. Analysis of their structures reveals that the increase in activity mainly occurs through the destabilization of the active site loop involved in substrate binding, and it has been observed that the level of disorder correlates with the width of the enzyme specificity spectrum. This finding supports the idea that active site conformational flexibility is essential to the acquisition of broader substrate specificity. | |||
Rational engineering of a native hyperthermostable lactonase into a broad spectrum phosphotriesterase.,Jacquet P, Hiblot J, Daude D, Bergonzi C, Gotthard G, Armstrong N, Chabriere E, Elias M Sci Rep. 2017 Dec 1;7(1):16745. doi: 10.1038/s41598-017-16841-0. PMID:29196634<ref>PMID:29196634</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[ | </div> | ||
[[Category: | <div class="pdbe-citations 5w3w" style="background-color:#fffaf0;"></div> | ||
[[Category: | |||
[[Category: Bergonzi | ==See Also== | ||
[[Category: | *[[Serum Paraoxonase|Serum Paraoxonase]] | ||
[[Category: Daude | == References == | ||
[[Category: Elias | <references/> | ||
[[Category: Hiblot | __TOC__ | ||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Saccharolobus solfataricus]] | |||
[[Category: Bergonzi C]] | |||
[[Category: Chabriere E]] | |||
[[Category: Daude D]] | |||
[[Category: Elias M]] | |||
[[Category: Gotthard G]] | |||
[[Category: Hiblot J]] | |||
[[Category: Jacquet P]] | |||
Latest revision as of 17:06, 4 October 2023
Crystal structure of SsoPox AsD6 mutant (V27A-Y97W-L228M-W263M) - open formCrystal structure of SsoPox AsD6 mutant (V27A-Y97W-L228M-W263M) - open form
Structural highlights
FunctionPHP_SACS2 Has a low paraoxonase activity. Also active, but with a lower activity, against other organo-phosphorus insecticides such as Dursban, Coumaphos, pNP-butanoate or parathion.[1] Publication Abstract from PubMedThe redesign of enzyme active sites to alter their function or specificity is a difficult yet appealing challenge. Here we used a structure-based design approach to engineer the lactonase SsoPox from Sulfolobus solfataricus into a phosphotriesterase. The five best variants were characterized and their structure was solved. The most active variant, alphasD6 (V27A-Y97W-L228M-W263M) demonstrates a large increase in catalytic efficiencies over the wild-type enzyme, with increases of 2,210-fold, 163-fold, 58-fold, 16-fold against methyl-parathion, malathion, ethyl-paraoxon, and methyl-paraoxon, respectively. Interestingly, the best mutants are also capable of degrading fensulfothion, which is reported to be an inhibitor for the wild-type enzyme, as well as others that are not substrates of the starting template or previously reported W263 mutants. The broad specificity of these engineered variants makes them promising candidates for the bioremediation of organophosphorus compounds. Analysis of their structures reveals that the increase in activity mainly occurs through the destabilization of the active site loop involved in substrate binding, and it has been observed that the level of disorder correlates with the width of the enzyme specificity spectrum. This finding supports the idea that active site conformational flexibility is essential to the acquisition of broader substrate specificity. Rational engineering of a native hyperthermostable lactonase into a broad spectrum phosphotriesterase.,Jacquet P, Hiblot J, Daude D, Bergonzi C, Gotthard G, Armstrong N, Chabriere E, Elias M Sci Rep. 2017 Dec 1;7(1):16745. doi: 10.1038/s41598-017-16841-0. PMID:29196634[2] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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