文献类型：期刊文献

英文题名：Structural and functional characterization of ubiquitin variant inhibitors for the JAMM-family deubiquitinases STAMBP and STAMBPL1

作者：Guo, Yusong[1,2];Liu, Qi[3];Mallette, Evan[3];Caba, Cody[4];Hou, Feng[2];Fux, Julia[3];LaPlante, Gabriel[3];Dong, Aiping[2];Zhang, Qi[2];Zheng, Hui[5,6];Tong, Yufeng[2,4];Zhang, Wei[3,7]

机构：[1]Guangdong Ocean Univ, Fisheries Coll, Zhanjiang, Guangdong, Peoples R China;[2]Univ Toronto, Struct Genom Consortium, Toronto, ON, Canada;[3]Univ Guelph, Coll Biol Sci, Dept Mol & Cellular Biol, Guelph, ON, Canada;[4]Univ Windsor, Dept Chem & Biochem, Windsor, ON, Canada;[5]Soochow Univ, Inst Biol, Int Inst Infect & Immun, Jiangsu Key Lab Infect & Immun, Suzhou, Jiangsu, Peoples R China;[6]Soochow Univ, Inst Med Sci, Int Inst Infect & Immun, Jiangsu Key Lab Infect & Immun, Suzhou, Jiangsu, Peoples R China;[7]Canadian Inst Adv Res, CIFAR Azrieli Global Scholars Program, Toronto, ON, Canada

年份：2021

卷号：297

期号：4

外文期刊名：JOURNAL OF BIOLOGICAL CHEMISTRY

收录：SCI-EXPANDED(收录号：WOS:000713004000011)、、EI(收录号：20213910934766)、Scopus(收录号：2-s2.0-85115365959)、WOS

基金：This research was funded by NSERC Discovery Grants awarded to W. Z. (RGPIN-2019-05721) and Y. T. (RGPIN-2017-06520) . Y. G. is a visiting scholar sponsored by the Excellent Young Teachers program and Innovation Fund of Guangdong Ocean University, China. Q. L. is supported by an OGS scholarship. E. M. was a MITACS Industrial Postdoctoral Fellow with funding provided by Mitacs and ProteinQure Inc. W. Z. is currently a CIFAR Azrieli Global Scholar in the Humans & The Microbiome Program. W. Z. is also a recipient of the Cancer Research Society/BMO Bank of Montreal Scholarship for the Next Generation of Scientists.

语种：英文

外文关键词：Amino acids - Biochemical engineering - Molecules - Proteins

外文摘要：Ubiquitination is a crucial posttranslational protein modifi- cation involved in a myriad of biological pathways. This modification is reversed by deubiquitinases (DUBs) that deconjugate the single ubiquitin (Ub) moiety or poly-Ub chains from substrates. In the past decade, tremendous efforts have been focused on targeting DUBs for drug discovery. However, most chemical compounds with inhibitory activity for DUBs suffer from mild potency and low selectivity. To overcome these obstacles, we developed a phage display-based protein engineering strategy for generating Ub variant (UbV) inhibitors, which was previously successfully applied to the Ubspecific protease (USP) family of cysteine proteases. In this work, we leveraged the UbV platform to selectively target STAMBP, a member of the JAB1/MPN/MOV34 (JAMM) metalloprotease family of DUB enzymes. We identified two UbVs (UbV(SP.1) and UbV(SP.3)) that bind to STAMBP with high affinity but differ in their selectivity for the closely related paralog STAMBPL1. We determined the STAMBPL1-UbV(SP.1) complex structure by X-ray crystallography, revealing hotspots of the JAMM-UbV interaction. Finally, we show that UbV(SP.1) and UbV(SP.3) are potent inhibitors of STAMBP isopeptidase activity, far exceeding the reported small-molecule inhibitor BC-1471. This work demonstrates that UbV technology is suitable to develop molecules as tools to target metalloproteases, which can be used to further understand the cellular function of JAMM family DUBs.