Mechanism of the Rpn13-induced activation of Uch37

doi:10.1007/s13238-014-0046-z

	Mechanism of the Rpn13-induced activation of Uch37
	Jiao, Lianying 1; Ouyang, Songying 1; Shaw, Neil 1; Song, Gaojie2 ; Feng, Yingang 3; Niu, Fengfeng 1; Qiu, Weicheng 1; Zhu, Hongtao 1; Hung, Li-Wei 4; Zuo, Xiaobing 5; Shtykova, V. Eleonora 6; Zhu, Ping 1; Dong, Yu-Hui 7; Xu, Ruxiang 8; Liu, Zhi-Jie 1,3
	2014-08
发表期刊	PROTEIN & CELL
ISSN	1674-800X
卷号	5 期号:8 页码:616-630
发表状态	已发表
DOI	10.1007/s13238-014-0046-z
摘要	Uch37 is a de-ubiquitinating enzyme that is activated by Rpn13 and involved in the proteasomal degradation of proteins. The full-length Uch37 was shown to exhibit low iso-peptidase activity and is thought to be auto-inhibited. Structural comparisons revealed that within a homo-dimer of Uch37, each of the catalytic domains was blocking the other's ubiquitin (Ub)-binding site. This blockage likely prevented Ub from entering the active site of Uch37 and might form the basis of auto-inhibition. To understand the mode of auto-inhibition clearly and shed light on the activation mechanism of Uch37 by Rpn13, we investigated the Uch37-Rpn13 complex using a combination of mutagenesis, biochemical, NMR, and small-angle X-ray scattering (SAXS) techniques. Our results also proved that Uch37 oligomerized in solution and had very low activity against the fluorogenic substrate ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) of de-ubiquitinating enzymes. Uch37 Delta(Hb,Hc,KEKE), a truncation removal of the C-terminal extension region (residues 256-329) converted oligomeric Uch37 into a monomeric form that exhibited iso-peptidase activity comparable to that of a truncation-containing the Uch37 catalytic domain only. We also demonstrated that Rpn13C (Rpn13 residues 270-407) could disrupt the oligomerization of Uch37 by sequestering Uch37 and forming a Uch37-Rpn13 complex. Uch37 was activated in such a complex, exhibiting 12-fold-higher activity than Uch37 alone. Time-resolved SAXS (TR-SAXS) and FRET experiments supported the proposed mode of auto-inhibition and the activation mechanism of Uch37 by Rpn13. Rpn13 activated Uch37 by forming a 1:1 stoichiometric complex in which the active site of Uch37 was accessible to Ub.
关键词	Uch37-Rpn13 complex de-ubiquitination SAXS analysis oligomerization iso-peptidase
收录类别	SCI ; CSCD
语种	英语
资助项目	National Institute of General Medical Sciences from the National Institutes of Health[9 P41 GM103622-17]
WOS研究方向	Cell Biology
WOS类目	Cell Biology
WOS记录号	WOS:000340566500005
CSCD记录号	CSCD:5242655
出版者	SPRINGEROPEN
WOS关键词	DEUBIQUITINATING ENZYME UCH37 ; SITE CROSSOVER LOOP ; PROTEASOME SUBUNIT ; CRYSTAL-STRUCTURE ; PROTEINS ; SYSTEM ; PROGRAM ; ROLES ; SPECIFICITY ; REVEALS
原始文献类型	Article
引用统计
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/1066
专题	iHuman研究所_PI研究组_刘志杰组
通讯作者	Xu, Ruxiang
作者单位	1.Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China 2.ShanghaiTech Univ, iHuman Inst, Shanghai 201210, Peoples R China 3.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Shandong Prov Key Lab Energy Genet, Qingdao 266101, Peoples R China 4.Los Alamos Natl Lab, Div Phys, Los Alamos, NM 87545 USA 5.Argonne Natl Lab, Xray Sci Div, Argonne, IL 60439 USA 6.Russian Acad Sci, Inst Crystallog, Moscow 117333, Russia 7.Chinese Acad Sci, Inst High Energy Phys, Ctr Multidisciplinary Res, Beijing 100049, Peoples R China 8.Mil Gen Hosp Beijing PLA, Dept Neurosurg, Beijing 100700, Peoples R China
推荐引用方式 GB/T 7714	Jiao, Lianying,Ouyang, Songying,Shaw, Neil,et al. Mechanism of the Rpn13-induced activation of Uch37[J]. PROTEIN & CELL,2014,5(8):616-630.
APA	Jiao, Lianying.,Ouyang, Songying.,Shaw, Neil.,Song, Gaojie.,Feng, Yingang.,...&Liu, Zhi-Jie.(2014).Mechanism of the Rpn13-induced activation of Uch37.PROTEIN & CELL,5(8),616-630.
MLA	Jiao, Lianying,et al."Mechanism of the Rpn13-induced activation of Uch37".PROTEIN & CELL 5.8(2014):616-630.