Expression, purification and characterization of the second DUSP domain of deubiquitinase USP20/VDU2

doi:10.1016/j.pep.2021.105836

	Expression, purification and characterization of the second DUSP domain of deubiquitinase USP20/VDU2
	Hu, Shuwei 1; Chen, Xiaoyu 1,2; Zhu, Mengying 1,2; Hu, Huifang 1,2; Zhou, Chen 1; Shi, Li 1,2; Zhang, Naixia 1,2
刊名	PROTEIN EXPRESSION AND PURIFICATION
	2021-05-01
卷号	181 页码:6
关键词	Deubiquitinase USP20 DUSP domain NMR
ISSN号	1046-5928
DOI	10.1016/j.pep.2021.105836
通讯作者	Shi, Li(shili@simm.ac.cn) ; Zhang, Naixia(nxzhang@simm.ac.cn)
英文摘要	Deubiquitinase USP20/VDU2 (VHL-interacting Deubiquitinating Enzyme 2) has been proved to play vital roles in multiple cellular processes by controlling the life-span of substrate proteins including hypoxia-inducible factor HIF1 alpha, beta 2-adrenergic receptor, and type 2 iodothyronine deiodinase etc. USP20 contains four distinct structural domains, which include the N-terminal zinc-finger ubiquitin binding domain (ZnF-UBP), the catalytic domain (USP domain), and two tandem DUSP domains (DUSP1 and DUSP2). Here in this study, we report the setting up of the production approach for USP20 DUSP2, and the NMR characterization of the produced target protein. With the assistance of GB1 tag and glycerol, both the solubility and stability of USP20 DUSP2 are significantly enhanced. And by using the optimized protein production procedure, monomeric and stable N-15, C-13-labeled USP20 DUSP2 sample for NMR data acquisition was obtained. The secondary structural elements of USP20 DUSP2 were then revealed by the analysis of recorded NMR spectra, and USP20 DUSP2 forms an AB3 fold in solution. The production protocol and NMR characterization results reported in this manuscript could be utilized in the extended structural and functional studies of USP20 DUSP2.
资助项目	National Natural Science Foundation of China[32000890] ; National Natural Science Foundation of China[21977105] ; National Natural Science Foundation of China[21778061]
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000621214000007
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/295609]
专题	中国科学院上海药物研究所
通讯作者	Shi, Li; Zhang, Naixia
作者单位	1.Chinese Acad Sci, Shanghai Inst Mat Med, Dept Analyt Chem, 555 Zu Chong Zhi Rd, Shanghai 201203, Peoples R China 2.Univ Chinese Acad Sci, 19A Yuquan Rd, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Hu, Shuwei,Chen, Xiaoyu,Zhu, Mengying,et al. Expression, purification and characterization of the second DUSP domain of deubiquitinase USP20/VDU2[J]. PROTEIN EXPRESSION AND PURIFICATION,2021,181:6.
APA	Hu, Shuwei.,Chen, Xiaoyu.,Zhu, Mengying.,Hu, Huifang.,Zhou, Chen.,...&Zhang, Naixia.(2021).Expression, purification and characterization of the second DUSP domain of deubiquitinase USP20/VDU2.PROTEIN EXPRESSION AND PURIFICATION,181,6.
MLA	Hu, Shuwei,et al."Expression, purification and characterization of the second DUSP domain of deubiquitinase USP20/VDU2".PROTEIN EXPRESSION AND PURIFICATION 181(2021):6.