Crystal structure and enzymatic characterization of thymidylate synthase X from Helicobacter pylori strain SS1

doi:10.1002/pro.668

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	Crystal structure and enzymatic characterization of thymidylate synthase X from Helicobacter pylori strain SS1
	Wang, Kuifeng; Wang, Qi; Chen, Jing; Chen, Lili; Jiang, Hualiang; Shen, Xu
刊名	PROTEIN SCIENCE
	2011-08
卷号	20 期号:8 页码:1398-1410
关键词	thymidylate synthase Helicobacter pylori thymidylate synthase ThyA
ISSN号	0961-8368
DOI	10.1002/pro.668
文献子类	Article
英文摘要	Thymidylate synthase X (ThyX) catalyzes the methylation of dUMP to form dTMP in bacterial life cycle and is regarded as a promising target for antibiotics discovery. Helicobacter pylori is a human pathogen associated with a number of human diseases. Here, we cloned and purified the ThyX enzyme from H. pylori SS1 strain (HpThyX). The recombinant HpThyX was discovered to exhibit the maximum activity at pH 8.5, and K-m values of the two substrates dUMP and CH(2)H(4)folate were determined to be 15.3 +/- 1.25 mu M and 0.35 +/- 0.18 mM, respectively. The analyzed crystal structure of HpThyX with the cofactor FAD and the substrate dUMP (at 2.31 angstrom) revealed that the enzyme was a tetramer bound to four dUMP and four FAD molecules. Different from the catalytic feature of the classical thymidylate synthase (ThyA), N5 atom of the FAD functioned as a nucleophile in the catalytic reaction instead of Ser84 and Ser85 residues. Our current work is expected to help better understand the structural and enzymatic features of HpThyX thus further providing valuable information for anti-H. pylori inhibitor discovery.
资助项目	State Key Program of Basic Research of China[2010CB912501] ; State Key Program of Basic Research of China[2009CB918502] ; National Natural Science Foundation of China[30925040] ; National Natural Science Foundation of China[10979072] ; Foundation of Chinese Academy of Sciences[KSCX2-YW-R-168] ; Foundation of Chinese Academy of Sciences[KSCX1-YW-02-2]
WOS关键词	CATALYTIC MECHANISM ; COMPLEMENTING PROTEIN ; FUNCTIONAL-ANALYSIS ; FLAVIN ; THYX ; BIOSYNTHESIS ; ERADICATION ; CLONING
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	WILEY
WOS记录号	WOS:000293137300008
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/278460]
专题	药物安全性评价中心中科院受体结构与功能重点实验室新药研究国家重点实验室药理学第三研究室
通讯作者	Chen, Jing
作者单位	Chinese Acad Sci, State Key Lab Drug Res, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Wang, Kuifeng,Wang, Qi,Chen, Jing,et al. Crystal structure and enzymatic characterization of thymidylate synthase X from Helicobacter pylori strain SS1[J]. PROTEIN SCIENCE,2011,20(8):1398-1410.
APA	Wang, Kuifeng,Wang, Qi,Chen, Jing,Chen, Lili,Jiang, Hualiang,&Shen, Xu.(2011).Crystal structure and enzymatic characterization of thymidylate synthase X from Helicobacter pylori strain SS1.PROTEIN SCIENCE,20(8),1398-1410.
MLA	Wang, Kuifeng,et al."Crystal structure and enzymatic characterization of thymidylate synthase X from Helicobacter pylori strain SS1".PROTEIN SCIENCE 20.8(2011):1398-1410.