Structure and allosteric regulation of human NAD-dependent isocitrate dehydrogenase

doi:10.1038/s41421-020-00220-7

	Structure and allosteric regulation of human NAD-dependent isocitrate dehydrogenase
	Sun, Pengkai 1; Liu, Yan2 ; Ma, Tengfei 1; Ding, Jianping1,2,3
	2020-12-22
发表期刊	CELL DISCOVERY (IF:13.0[JCR-2023],14.8[5-Year])
卷号	6 期号:1
发表状态	已发表
DOI	10.1038/s41421-020-00220-7
摘要	Human NAD-dependent isocitrate dehydrogenase or HsIDH3 catalyzes the decarboxylation of isocitrate into alpha-ketoglutarate in the TCA cycle. HsIDH3 exists and functions as a heterooctamer composed of the alpha beta and alpha gamma heterodimers, and is regulated allosterically and/or competitively by numerous metabolites including CIT, ADP, ATP, and NADH. In this work, we report the crystal structure of HsIDH3 containing a beta mutant in apo form. In the HsIDH3 structure, the alpha beta and alpha gamma heterodimers form the alpha(2)beta gamma heterotetramer via their clasp domains, and two alpha(2)beta gamma heterotetramers form the (alpha(2)beta gamma)(2) heterooctamer through insertion of the N-terminus of the gamma subunit of one heterotetramer into the back cleft of the beta subunit of the other heterotetramer. The functional roles of the key residues at the allosteric site, the pseudo allosteric site, the heterodimer and heterodimer-heterodimer interfaces, and the N-terminal of the gamma subunit are validated by mutagenesis and kinetic studies. Our structural and biochemical data together demonstrate that the allosteric site plays an important role but the pseudo allosteric site plays no role in the allosteric activation of the enzyme; the activation signal from the allosteric site is transmitted to the active sites of both alpha beta and alpha gamma heterodimers via the clasp domains; and the N-terminal of the gamma subunit plays a critical role in the formation of the heterooctamer to ensure the optimal activity of the enzyme. These findings reveal the molecular mechanism of the assembly and allosteric regulation of HsIDH3.
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收录类别	SCI ; SCIE
语种	英语
资助项目	National Natural Science Foundation of China[31870723][31530013]
WOS研究方向	Cell Biology
WOS类目	Cell Biology
WOS记录号	WOS:000602688500001
出版者	SPRINGERNATURE
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/124922
专题	生命科学与技术学院_硕士生生命科学与技术学院_特聘教授组_丁建平组
通讯作者	Ding, Jianping
作者单位	1.Chinese Acad Sci, Univ Chinese Acad Sci, Ctr Excellence Mol Cell Sci, State Key Lab Mol Biol,Shanghai Inst Biochem & Ce, 320 Yueyang Rd, Shanghai 200031, Peoples R China 2.ShanghaiTech Univ, Sch Life Sci & Technol, 393 Huaxia Zhong Rd, Shanghai 201210, Peoples R China 3.Univ Chinese Acad Sci, Sch Life Sci, Hangzhou Inst Adv Study, 1 Xiangshan Rd, Hangzhou 310024, Zhejiang, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Sun, Pengkai,Liu, Yan,Ma, Tengfei,et al. Structure and allosteric regulation of human NAD-dependent isocitrate dehydrogenase[J]. CELL DISCOVERY,2020,6(1).
APA	Sun, Pengkai,Liu, Yan,Ma, Tengfei,&Ding, Jianping.(2020).Structure and allosteric regulation of human NAD-dependent isocitrate dehydrogenase.CELL DISCOVERY,6(1).
MLA	Sun, Pengkai,et al."Structure and allosteric regulation of human NAD-dependent isocitrate dehydrogenase".CELL DISCOVERY 6.1(2020).