Structural insights into the putative bacterial acetylcholinesterase ChoE and its substrate inhibition mechanism

doi:10.1074/jbc.ra119.011809

	Structural insights into the putative bacterial acetylcholinesterase ChoE and its substrate inhibition mechanism
	Pham, Van Dung 1,2,3; To, Tuan Anh 1,2,3; Gagné-Thivierge, Cynthia 1,2,4; Couture, Manon 1,2,3; Lagüe, Patrick 1,2,3; Yao, Deqiang5 ; Picard, Marie-Ève 1,2,3; Lortie, Louis-André 1; Attéré, Sabrina A.1,2,4; Zhu, Xiaojun 1,2,3; Levesque, Roger C.2; Charette, Steve J.1,2,4; Shi, Rong 1,2,3
	2020-06-26
发表期刊	JOURNAL OF BIOLOGICAL CHEMISTRY (IF:4.0[JCR-2023],4.4[5-Year])
ISSN	0021-9258
EISSN	1083-351X
卷号	295 期号:26 页码:8708-8724
发表状态	已发表
DOI	10.1074/jbc.ra119.011809
摘要	Mammalian acetylcholinesterase (AChE) is well-studied, being important in both cholinergic brain synapses and the peripheral nervous systems and also a key drug target for many diseases. In contrast, little is known about the structures and molecular mechanism of prokaryotic acetylcholinesterases. We report here the structural and biochemical characterization of ChoE, a putative bacterial acetylcholinesterase from Pseudomonas aeruginosa. Analysis of WT and mutant strains indicated that ChoE is indispensable for P. aeruginosa growth with acetylcholine as the sole carbon and nitrogen source. The crystal structure of ChoE at 1.35 Å resolution revealed that this enzyme adopts a typical fold of the SGNH hydrolase family. Although ChoE and eukaryotic AChEs catalyze the same reaction, their overall structures bear no similarities constituting an interesting example of convergent evolution. Among Ser-38, Asp-285, and His-288 of the catalytic triad residues, only Asp-285 was not essential for ChoE activity. Combined with kinetic analyses of WT and mutant proteins, multiple crystal structures of ChoE complexed with substrates, products, or reaction intermediate revealed the structural determinants for substrate recognition, snapshots of the various catalytic steps, and the molecular basis of substrate inhibition at high substrate concentrations. Our results indicate that substrate inhibition in ChoE is due to acetate release being blocked by the binding of a substrate molecule in a nonproductive mode. Because of the distinct overall folds and significant differences of the active site between ChoE and eukaryotic AChEs, these structures will serve as a prototype for other prokaryotic acetylcholinesterases. © 2020 American Society for Biochemistry and Molecular Biology Inc.. All rights reserved.
关键词	Mammals Reaction intermediates Substrates Biochemical characterization Convergent evolution High substrate concentrations Peripheral nervous system Pseudomonas aeruginosa Sgnh hydrolase families Structural determinants Substrate recognition acetylcholinesterase crystal structure enzyme catalysis catalytic triad substrate inhibition nonproductive binding convergent evolution acyl-enzyme serine esterase acetylcholinesterase (AChE)
收录类别	EI ; SCI ; SCIE
语种	英语
资助项目	Fonds de Recherche du Quebec-Nature et technologies (FRQNT)[183530]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000546296800008
出版者	American Society for Biochemistry and Molecular Biology Inc.
EI入藏号	20210109727760
EI主题词	Crystal structure
EI分类号	804 Chemical Products Generally ; 933.1.1 Crystal Lattice
WOS关键词	PSEUDOMONAS-AERUGINOSA ; CRYSTAL-STRUCTURE ; BACILLUS-SUBTILIS ; CATALYTIC SERINE ; CHOLINESTERASE ; ESTERASE ; ENZYME ; RESIDUES ; BINDING ; FAMILY
原始文献类型	Journal article (JA)
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/251747
专题	iHuman研究所_科学装置(X)_膜蛋白同步辐射线站
作者单位	1.Département de Biochimie, de Microbiologie et de Bio-informatique, Université Laval, Québec, Canada; 2.Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, Québec, Canada; 3.PROTEO, Québec Network for Research on Protein Function, Engineering, and Applications, Québec, Canada; 4.Centre de Recherche, Institut Universitaire de Cardiologie et de Pneumologie de Québec, Hôpital Laval, Québec, Canada; 5.IHuman Institute, ShanghaiTech University, Shanghai, China
推荐引用方式 GB/T 7714	Pham, Van Dung,To, Tuan Anh,Gagné-Thivierge, Cynthia,et al. Structural insights into the putative bacterial acetylcholinesterase ChoE and its substrate inhibition mechanism[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2020,295(26):8708-8724.
APA	Pham, Van Dung.,To, Tuan Anh.,Gagné-Thivierge, Cynthia.,Couture, Manon.,Lagüe, Patrick.,...&Shi, Rong.(2020).Structural insights into the putative bacterial acetylcholinesterase ChoE and its substrate inhibition mechanism.JOURNAL OF BIOLOGICAL CHEMISTRY,295(26),8708-8724.
MLA	Pham, Van Dung,et al."Structural insights into the putative bacterial acetylcholinesterase ChoE and its substrate inhibition mechanism".JOURNAL OF BIOLOGICAL CHEMISTRY 295.26(2020):8708-8724.