A fluorescence assay for elucidating the substrate specificities of deubiquitinating enzymes

	A fluorescence assay for elucidating the substrate specificities of deubiquitinating enzymes
	Yin, ST; Huang, H; Zhang, YH; Zhou, ZR; Song, AX; Hong, FS; Hu, HY
刊名	BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
	2011
卷号	416 期号:1 页码:76-79
关键词	Ubiquitin C-terminal hydrolase Substrate specificity Ni(2+)-NTA affinity Fluorescence Deubiquitinating enzyme
通讯作者	Hu, HY (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Shanghai 200031, Peoples R China.,hyhu@sibs.ac.cn
英文摘要	Ubiquitin C-terminal hydrolases (UCHs) are a representative family of deubiquitinating enzymes (DUBs), which specifically cleave ubiquitin,(Ub) chains or extensions. Here we present a convenient method for characterizing the substrate specificities of various UCHs by fluorescently mutated Ub-fusion proteins (Ub(F45W)-Xaa) and di-ubiquitin chains (Ub(F45W)-diUb). After removal of the intact substrate by Ni(2+)-NTA affinity, the enzymatic activities of UCHs were quantitatively determined by recording fluorescence of the Ub(F45W) product. The results show that three UCHs, i.e. UCH-L1. UCH-L3 and UCH37/UCH-L5, are distinct in their substrate specificities for the Ub-fusions and diUb chains. This assay method may also be applied to study the enzymatic activities and substrate specificities of other DUBs. (C) 2011 Elsevier Inc. All rights reserved.
学科主题	Biochemistry & Molecular Biology; Biophysics
类目[WOS]	Biochemistry & Molecular Biology ; Biophysics
关键词[WOS]	RESONANCE-ENERGY-TRANSFER ; UBIQUITIN ; PROTEIN ; UCH37 ; HYDROLASES
收录类别	SCI
语种	英语
WOS记录号	WOS:000298222100013
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/897]
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	Yin, ST,Huang, H,Zhang, YH,et al. A fluorescence assay for elucidating the substrate specificities of deubiquitinating enzymes[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2011,416(1):76-79.
APA	Yin, ST.,Huang, H.,Zhang, YH.,Zhou, ZR.,Song, AX.,...&Hu, HY.(2011).A fluorescence assay for elucidating the substrate specificities of deubiquitinating enzymes.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,416(1),76-79.
MLA	Yin, ST,et al."A fluorescence assay for elucidating the substrate specificities of deubiquitinating enzymes".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 416.1(2011):76-79.