X-ray crystallographic and enzymatic analyses of shikimate dehydrogenase from Staphylococcus epidermidis

	X-ray crystallographic and enzymatic analyses of shikimate dehydrogenase from Staphylococcus epidermidis
	Han, C; Hu, TC; Wu, DL; Qu, S; Zhou, JH; Ding, JP; Shen, X; Qu, D; Jiang, HL
刊名	FEBS JOURNAL
	2009
卷号	276 期号:4 页码:1125-1139
关键词	crystal structure shikimate dehydrogenase shikimate pathway site-directed mutagenesis Staphylococcus epidermidis
通讯作者	Qu, D (reprint author), Fudan Univ, Shanghai Med Coll, Inst Med Microbiol, Inst Biomed Sci, Shanghai 200032, Peoples R China.,xshen@mail.shcnc.ac.cn ; dqu@shmu.edu.cn
英文摘要	Shikimate dehydrogenase (SDH) catalyzes the NADPH-dependent reduction of 3-dehydroshikimate to shikimate in the shikimate pathway. In this study, we determined the kinetic properties and crystal structures of Staphylococcus epidermidis SDH (SeSDH) both in its ligand-free form and in complex with shikimate. SeSDH has a k(cat) of 22.8 s(-1) and a K(m) of 73 mu m towards shikimate, and a K(m) of 100 mu m towards NADP. The overall folding of SeSDH comprises the N-terminal alpha/beta domain for substrate binding and the C-terminal Rossmann fold for NADP binding. The active site is within a large groove between the two domains. Residue Tyr211, normally regarded as important for substrate binding, does not interact with shikimate in the binary SeSDH-shikimate complex structure. However, the Y211F mutation leads to a significant decrease in k(cat) and a minor increase in the K(m) for shikimate. The results indicate that the main function of Tyr211 may be to stabilize the catalytic intermediate during catalysis. The NADP-binding domain of SeSDH is less conserved. The usually long helix specifically recognizing the adenine ribose phosphate is substituted with a short 3(10) helix in the NADP-binding domain. Moreover, the interdomain angle of SeSDH is the widest among all known SDH structures, indicating an inactive 'open' state of the SeSDH structure. Thus, a 'closing' process might occur upon NADP binding to bring the cofactor close to the substrate for catalysis.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	AMINO-ACID BIOSYNTHESIS ; CRYSTAL-STRUCTURE ; MYCOBACTERIUM-TUBERCULOSIS ; 5-DEHYDROGENASE ; PATHWAY ; AROE ; INSIGHTS ; INFECTIONS ; MECHANISM ; DENSITY
收录类别	SCI
语种	英语
WOS记录号	WOS:000262666900021
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/1183]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Han, C,Hu, TC,Wu, DL,et al. X-ray crystallographic and enzymatic analyses of shikimate dehydrogenase from Staphylococcus epidermidis[J]. FEBS JOURNAL,2009,276(4):1125-1139.
APA	Han, C.,Hu, TC.,Wu, DL.,Qu, S.,Zhou, JH.,...&Jiang, HL.(2009).X-ray crystallographic and enzymatic analyses of shikimate dehydrogenase from Staphylococcus epidermidis.FEBS JOURNAL,276(4),1125-1139.
MLA	Han, C,et al."X-ray crystallographic and enzymatic analyses of shikimate dehydrogenase from Staphylococcus epidermidis".FEBS JOURNAL 276.4(2009):1125-1139.