File Download
  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Crystal Structure of the Thioesterification Conformation of Bacillus Subtilis O-succinylbenzoyl-CoA Synthetase Reveals a Distinct Substrate-binding Mode

TitleCrystal Structure of the Thioesterification Conformation of Bacillus Subtilis O-succinylbenzoyl-CoA Synthetase Reveals a Distinct Substrate-binding Mode
Authors
Issue Date2017
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal of Biological Chemistry, 2017, v. 292 n. 29, p. 12296-12310 How to Cite?
Abstracto-Succinylbenzoyl-CoA (OSB-CoA) synthetase (MenE) is an essential enzyme in bacterial vitamin K biosynthesis and an important target in the development of new antibiotics. It is a member of the adenylating enzymes (ANL) family, which reconfigure their active site in two different active conformations, one for the adenylation half-reaction and the other for a thioesterification half-reaction, in a domain-alternation catalytic mechanism. Although several aspects of the adenylating mechanism in MenE have recently been uncovered, its thioesterification conformation remains elusive. Here, using a catalytically competent Bacillus subtilis mutant protein complexed with an OSB-CoA analogue, we determined MenE high-resolution structures to 1.76 and 1.90 Å resolution in a thioester-forming conformation. By comparison with the adenylation conformation, we found that MenE’s C-domain rotates around the Ser-384 hinge by 139.5° during domain-alternation catalysis. The structures also revealed a thioesterification active site specifically conserved among MenE orthologues and a substrate-binding mode distinct from those of many other acyl/aryl-CoA synthetases. Of note, using site-directed mutagenesis, we identified several residues that specifically contribute to the thioesterification half-reaction without affecting the adenylation half-reaction. Moreover, we observed a substantial movement of the activated succinyl group in the thioesterification half-reaction. These findings provide new insights into the domain-alternation catalysis of a bacterial enzyme essential for vitamin K biosynthesis and of its adenylating homologues in the ANL enzyme family.
Persistent Identifierhttp://hdl.handle.net/10722/252185
ISSN
2017 Impact Factor: 4.011
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorChen, Y-
dc.contributor.authorLi, T-
dc.contributor.authorLin, X-
dc.contributor.authorLi, X-
dc.contributor.authorLi, XD-
dc.contributor.authorGuo, Z-
dc.date.accessioned2018-04-12T01:16:17Z-
dc.date.available2018-04-12T01:16:17Z-
dc.date.issued2017-
dc.identifier.citationJournal of Biological Chemistry, 2017, v. 292 n. 29, p. 12296-12310-
dc.identifier.issn0021-9258-
dc.identifier.urihttp://hdl.handle.net/10722/252185-
dc.description.abstracto-Succinylbenzoyl-CoA (OSB-CoA) synthetase (MenE) is an essential enzyme in bacterial vitamin K biosynthesis and an important target in the development of new antibiotics. It is a member of the adenylating enzymes (ANL) family, which reconfigure their active site in two different active conformations, one for the adenylation half-reaction and the other for a thioesterification half-reaction, in a domain-alternation catalytic mechanism. Although several aspects of the adenylating mechanism in MenE have recently been uncovered, its thioesterification conformation remains elusive. Here, using a catalytically competent Bacillus subtilis mutant protein complexed with an OSB-CoA analogue, we determined MenE high-resolution structures to 1.76 and 1.90 Å resolution in a thioester-forming conformation. By comparison with the adenylation conformation, we found that MenE’s C-domain rotates around the Ser-384 hinge by 139.5° during domain-alternation catalysis. The structures also revealed a thioesterification active site specifically conserved among MenE orthologues and a substrate-binding mode distinct from those of many other acyl/aryl-CoA synthetases. Of note, using site-directed mutagenesis, we identified several residues that specifically contribute to the thioesterification half-reaction without affecting the adenylation half-reaction. Moreover, we observed a substantial movement of the activated succinyl group in the thioesterification half-reaction. These findings provide new insights into the domain-alternation catalysis of a bacterial enzyme essential for vitamin K biosynthesis and of its adenylating homologues in the ANL enzyme family.-
dc.languageeng-
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/-
dc.relation.ispartofJournal of Biological Chemistry-
dc.rightsJournal of Biological Chemistry. Copyright © American Society for Biochemistry and Molecular Biology, Inc.-
dc.rightsThis research was originally published in [Journal of Biological Chemistry]. Yaozong Chen, Tin Lok Li, Xingbang Lin, Xin Li, Xiang David Li and Zhihong Guo. Crystal Structure of the Thioesterification Conformation of Bacillus Subtilis O-succinylbenzoyl-CoA Synthetase Reveals a Distinct Substrate-binding Mode. Journal of Biological Chemistry, 2017, v. 292 n. 29, p. 12296-12310 © the American Society for Biochemistry and Molecular Biology-
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.titleCrystal Structure of the Thioesterification Conformation of Bacillus Subtilis O-succinylbenzoyl-CoA Synthetase Reveals a Distinct Substrate-binding Mode-
dc.typeArticle-
dc.identifier.emailLi, X: lx418@hku.hk-
dc.identifier.emailLi, XD: xiangli@hku.hk-
dc.identifier.authorityLi, XD=rp01562-
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1074/jbc.M117.790410-
dc.identifier.hkuros284607-
dc.identifier.volume292-
dc.identifier.issue29-
dc.identifier.spage12296-
dc.identifier.epage12310-
dc.identifier.isiWOS:000406053300025-
dc.publisher.placeUnited States-

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats