Structures of trans-2-enoyl-00A reductases from Clostridium acelobutylicum and Treponema denticola: insights into the substrate specificity and the catalytic mechanism

	Structures of trans-2-enoyl-00A reductases from Clostridium acelobutylicum and Treponema denticola: insights into the substrate specificity and the catalytic mechanism
	Hu, K; Zhao, M; Zhang, TL; Zha, MW; Zhong, C; Jiang, Y; Ding, JP
刊名	BIOCHEMICAL JOURNAL
	2013
卷号	449 期号:1 页码:79-89
关键词	biofuel catalytic mechanism crystal structure reductase substrate specificity synthetic biology
通讯作者	Jiang, Y (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, Key Lab Synthet Biol, 300 Feng Lin Rd, Shanghai 200031, Peoples R China.,yjiang@cibt.ac.cn ; jpding@sibs.ac.cn
英文摘要	TERs (trans-2-enoyl-CoA reductases; EC 1.3.1.44), which specifically catalyse the reduction of crotonyl-CoA to butyryl-CoA using NADH as cofactor, have recently been applied in the design of robust synthetic pathways to produce butan-1-ol as a biofuel. We report in the present paper the characterization of a CaTER (a TER homologue in Clostridium acetobutylicum), the structures of CaTER in apo form and in complexes with NADH and NAD(+) and the structure of TdTER (Treponema denticola TER) in complex with NAD(+). Structural and sequence comparisons show that CaTER and TdTER share approximately 45% overall sequence identity and high structural similarities with the FabV class enoyl-acyl carrier protein reductases in the bacterial fatty acid synthesis pathway, suggesting that both types of enzymes belong to the same family. CaTER and TdTER function as monomers and consist of a cofactor-binding domain and a substrate-binding domain with the catalytic active site located at the interface of the two domains. Structural analyses of CaTER together with mutagenesis and biochemical data indicate that the conserved Glu(75) determines the cofactor specificity, and the conserved Tyr(225), Tyr(235) and Lys(244) play critical roles in catalysis. Upon cofactor binding, the substrate-binding loop changes from an open conformation to a closed conformation, narrowing a hydrophobic channel to the catalytic site. A modelling study shows that the hydrophobic channel is optimal in both width and length for the binding of crotonyl-CoA. These results provide molecular bases for the high substrate specificity and the catalytic mechanism of TERs.
学科主题	Biochemistry & Molecular Biology
类目[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	ENOYL-ACP REDUCTASE ; MYCOBACTERIUM-TUBERCULOSIS ; ESCHERICHIA-COLI ; EUGLENA-GRACILIS ; BUTANOL ; CONSERVATION ; EXPRESSION ; PATHWAYS ; BIOFUEL ; COMPLEX
收录类别	SCI
语种	英语
WOS记录号	WOS:000312470800008
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/525]
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	Hu, K,Zhao, M,Zhang, TL,et al. Structures of trans-2-enoyl-00A reductases from Clostridium acelobutylicum and Treponema denticola: insights into the substrate specificity and the catalytic mechanism[J]. BIOCHEMICAL JOURNAL,2013,449(1):79-89.
APA	Hu, K.,Zhao, M.,Zhang, TL.,Zha, MW.,Zhong, C.,...&Ding, JP.(2013).Structures of trans-2-enoyl-00A reductases from Clostridium acelobutylicum and Treponema denticola: insights into the substrate specificity and the catalytic mechanism.BIOCHEMICAL JOURNAL,449(1),79-89.
MLA	Hu, K,et al."Structures of trans-2-enoyl-00A reductases from Clostridium acelobutylicum and Treponema denticola: insights into the substrate specificity and the catalytic mechanism".BIOCHEMICAL JOURNAL 449.1(2013):79-89.