A fluorescence assay for elucidating the substrate specificities of deubiquitinating enzymes
文献类型：期刊论文
| 作者 | Yin, ST; Huang, H; Zhang, YH; Zhou, ZR; Song, AX; Hong, FS; Hu, HY |
| 刊名 | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
 |
| 出版日期 | 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 |
| 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. |
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