Structure and Function of a C-C Bond Cleaving Oxygenase in Atypical Angucycline Biosynthesis

doi:10.1021/acschembio.6b00621

KMS > iHuman研究所 > 特聘教授组 > Raymond Stevens组

	Structure and Function of a C-C Bond Cleaving Oxygenase in Atypical Angucycline Biosynthesis
	Pan, Guohui 1; Gao, Xiaoqin 2,3; Fan, Keqiang 1; Liu, Junlin4 ; Meng, Bing4 ; Gao, Jinmin 1,3; Wang, Bin 1; Zhang, Chaobo 1,3; Han, Hui 1; Ai, Guomin 1; Chen, Yihua 1; Wu, Dong4 ; Liu, Zhi-Jie 2,5; Yang, Keqian 1
	2017-01
发表期刊	ACS CHEMICAL BIOLOGY (IF:3.5[JCR-2023],3.9[5-Year])
ISSN	1554-8929
卷号	12 期号:1 页码:142-152
发表状态	已发表
DOI	10.1021/acschembio.6b00621
摘要	C-C bond ring cleaving oxygenases represent a unique family of enzymes involved in the B ring cleavage reaction only observed in atypical angucycline biosynthesis. B ring cleavage is the key reaction leading to dramatic divergence in the final structures of atypical angucyclines. Here, we present the crystal structure of AlpJ, the first structure of this family of enzymes. AlpJ has been verified as the enzyme catalyzing C-C bond cleavage in kinamycin biosynthesis. The crystal structure of the AlpJ monomer resembles the dimeric structure of ferredoxin-like proteins. The N- and C-terminal halves of AlpJ are homologous, and both contain a putative hydrophobic substrate binding pocket in the "closed" and "open" conformations, respectively. Structural comparison of AlpJ with ActVA-Orf6 and protein ligand docking analysis suggest that the residues including Asn60, Trp64, and Trp181 are possibly involved in substrate recognition. Site-directed mutagenesis results supported our hypothesis, as mutation of these residues led to nearly a complete loss of the activity of AlpJ. Structural analysis also revealed that AlpJ possesses an intramolecular domain domain interface, where the residues His50 and Tyr178 form a hydrogen bond that probably stabilizes the three-dimensional structure of AlpJ. Site-directed mutagenesis showed that the two residues, His50 and Tyr178, were vital for the activity of AlpJ. Our findings shed light on the structure and catalytic mechanism of the AlpJ family of oxygenases, which presumably involves two active sites that might function in a cooperative manner.
收录类别	SCI
语种	英语
资助项目	Ministry of Science and Technology of China[2014CB910400] ; Ministry of Science and Technology of China[2015CB910104]
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000392569400016
出版者	AMER CHEMICAL SOC
WOS关键词	STREPTOMYCES-VENEZUELAE ISP5230 ; ANTITUMOR ANTIBIOTIC C-1027 ; GENE-CLUSTER ; JADOMYCIN-B ; HETEROLOGOUS EXPRESSION ; CLEAVAGE REACTION ; GILVOCARCIN-V ; CLONING ; MONOOXYGENASE ; FLUOSTATINS
原始文献类型	Article
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/1573
专题	iHuman研究所_特聘教授组_Raymond Stevens组免疫化学研究所_特聘教授组_抗体结构学实验室 iHuman研究所_公共科研平台_昆虫细胞培养平台 iHuman研究所_PI研究组_陶厚朝组
通讯作者	Wu, Dong; Liu, Zhi-Jie; Yang, Keqian
作者单位	1.Chinese Acad Sci, Inst Microbiol, State Key Lab Microbial Resources, Beijing 100101, Peoples R China 2.Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 4.ShanghaiTech Univ, iHuman Inst, Shanghai 201210, Peoples R China 5.Kunming Med Univ, Inst Mol & Clin Med, Kunming 650500, Peoples R China
通讯作者单位	iHuman研究所
推荐引用方式 GB/T 7714	Pan, Guohui,Gao, Xiaoqin,Fan, Keqiang,et al. Structure and Function of a C-C Bond Cleaving Oxygenase in Atypical Angucycline Biosynthesis[J]. ACS CHEMICAL BIOLOGY,2017,12(1):142-152.
APA	Pan, Guohui.,Gao, Xiaoqin.,Fan, Keqiang.,Liu, Junlin.,Meng, Bing.,...&Yang, Keqian.(2017).Structure and Function of a C-C Bond Cleaving Oxygenase in Atypical Angucycline Biosynthesis.ACS CHEMICAL BIOLOGY,12(1),142-152.
MLA	Pan, Guohui,et al."Structure and Function of a C-C Bond Cleaving Oxygenase in Atypical Angucycline Biosynthesis".ACS CHEMICAL BIOLOGY 12.1(2017):142-152.