File Download
Links for fulltext
(May Require Subscription)
- Publisher Website: 10.1074/jbc.M117.790410
- Scopus: eid_2-s2.0-85025173612
- WOS: WOS:000406053300025
- Find via
Supplementary
- Citations:
- Appears in Collections:
Article: Crystal Structure of the Thioesterification Conformation of Bacillus Subtilis O-succinylbenzoyl-CoA Synthetase Reveals a Distinct Substrate-binding Mode
Title | Crystal Structure of the Thioesterification Conformation of Bacillus Subtilis O-succinylbenzoyl-CoA Synthetase Reveals a Distinct Substrate-binding Mode |
---|---|
Authors | |
Issue Date | 2017 |
Publisher | American 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? |
Abstract | o-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 Identifier | http://hdl.handle.net/10722/252185 |
ISSN | 2020 Impact Factor: 5.157 2020 SCImago Journal Rankings: 2.361 |
ISI Accession Number ID |
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Chen, Y | - |
dc.contributor.author | Li, T | - |
dc.contributor.author | Lin, X | - |
dc.contributor.author | Li, X | - |
dc.contributor.author | Li, XD | - |
dc.contributor.author | Guo, Z | - |
dc.date.accessioned | 2018-04-12T01:16:17Z | - |
dc.date.available | 2018-04-12T01:16:17Z | - |
dc.date.issued | 2017 | - |
dc.identifier.citation | Journal of Biological Chemistry, 2017, v. 292 n. 29, p. 12296-12310 | - |
dc.identifier.issn | 0021-9258 | - |
dc.identifier.uri | http://hdl.handle.net/10722/252185 | - |
dc.description.abstract | o-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.language | eng | - |
dc.publisher | American Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/ | - |
dc.relation.ispartof | Journal of Biological Chemistry | - |
dc.rights | This research was originally published in the 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. J Biol Chem. 2017; 292:12296-12310. © the American Society for Biochemistry and Molecular Biology. | - |
dc.title | Crystal Structure of the Thioesterification Conformation of Bacillus Subtilis O-succinylbenzoyl-CoA Synthetase Reveals a Distinct Substrate-binding Mode | - |
dc.type | Article | - |
dc.identifier.email | Li, X: lx418@hku.hk | - |
dc.identifier.email | Li, XD: xiangli@hku.hk | - |
dc.identifier.authority | Li, XD=rp01562 | - |
dc.description.nature | published_or_final_version | - |
dc.identifier.doi | 10.1074/jbc.M117.790410 | - |
dc.identifier.scopus | eid_2-s2.0-85025173612 | - |
dc.identifier.hkuros | 284607 | - |
dc.identifier.volume | 292 | - |
dc.identifier.issue | 29 | - |
dc.identifier.spage | 12296 | - |
dc.identifier.epage | 12310 | - |
dc.identifier.isi | WOS:000406053300025 | - |
dc.publisher.place | United States | - |
dc.identifier.issnl | 0021-9258 | - |