文献类型：期刊文献

英文题名：Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10

作者：Zhang, Wei[1,2];Sartori, Maria A.[1,2];Makhnevych, Taras[1,2];Federowicz, Kelly E.[3];Dong, Xiaohui[4];Liu, Li[5];Nim, Satra[1,2];Dong, Aiping[6];Yang, Jingsong[7];Li, Yanjun[6];Haddad, Dania[1,2];Ernst, Andreas[1,2];Heerding, Dirk[3];Tong, Yufeng[6,8];Moffat, Jason[1,2];Sidhu, Sachdev S.[1,2]

机构：[1]Univ Toronto, Banting & Best Dept Med Res, Donnelly Ctr Cellular & Biomol Res, 160 Coll St, Toronto, ON M5S 3E1, Canada;[2]Univ Toronto, Dept Mol Genet, 160 Coll St, Toronto, ON M5S 3E1, Canada;[3]GlaxoSmithKline Inc, Oncol Res & Dev, Canc Epigenet DPU, 1250 S Collegeville Rd, Collegeville, PA 19426 USA;[4]Guangdong Ocean Univ, Fisheries Coll, Dept Aquaculture, Zhanjiang 524088, Guangdong, Peoples R China;[5]Guangdong Ocean Univ, Fisheries Coll, Dept Marine Biol, Zhanjiang 524025, Guangdong, Peoples R China;[6]Univ Toronto, Struct Genom Consortium, Toronto, ON M5G 1L7, Canada;[7]GlaxoSmithKline Inc, Oncol Res & Dev, Immunooncol Combinat DPU, 1250 S Collegeville Rd, Collegeville, PA 19426 USA;[8]Univ Toronto, Dept Pharmacol & Toxicol, Toronto, ON M5G 1L7, Canada

年份：2017

卷号：429

期号：22

起止页码：3546

外文期刊名：JOURNAL OF MOLECULAR BIOLOGY

收录：SCI-EXPANDED(收录号：WOS:000416188500013)、、Scopus(收录号：2-s2.0-85020850842)、WOS

基金：We are grateful to Dr. Bachir Affar for helpful discussions, and Mr. Jun Gu for technical support. We acknowledge the mass spectrometry technical contribution from the group of Dr. Jack Greenblatt. The Structural Genomics Consortium (SGC) is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada through Ontario Genomics Institute, Innovative Medicines Initiative (EU/EFPIA; ULTRA-DD grant no. 115766), Janssen, Merck & Co., Novartis Pharma AG, Ontario Ministry of Economic Development and Innovation, Pfizer, Sao Paulo Research Foundation-FAPESP, Takeda, and the Wellcome Trust. X-ray diffraction data were collected at Argonne National Laboratory, operated for the U.S. Department of Energy under contract DE-AC02-06CH11357. L.L and X.D. are visiting scholars sponsored by the National Natural Science Foundation of China. W.Z. is supported by the Mitacs Elevate program with funds from Mitacs Inc. and the Centre for Commercialization of Antibodies and Biologies (CCAB). This work is supported by a Canadian Cancer Society Research Institute (CCSRI) Innovation grant (#702861) and the Canadian Institutes for Health Research (CIHR) project grant (#0000303157) awarded to J.M. and S.S.S. and by a Genome Canada Disruptive Innovation in Genomics grant (OGI-119) awarded to S.S.S through Ontario Genomics.

语种：英文

外文关键词：USP7; USP10; p53; inhibitor design

外文摘要：Post-translational modification of the p53 signaling pathway plays an important role in cell cycle progression and stress-induced apoptosis. Indeed, a large body of work has shown that dysregulation of p53 and its E3 ligase MDM2 by the ubiquitin-proteasome system (UPS) promotes carcinogenesis and malignant transformation. Thus, drug discovery efforts have focused on the restoration of wild-type p53 activity or inactivation of oncogenic mutant p53 by targeted inhibition of UPS components, particularly key deubiquitinases (DUBs) of the ubiquitin-specific protease (USP) class. However, development of selective small-molecule USP inhibitors has been challenging, partly due to the highly conserved structural features of the catalytic sites across the class. To tackle this problem, we devised a protein engineering strategy for rational design of inhibitors for DUBs and other UPS proteins. We employed a phage-displayed ubiquitin variant (UbV) library to develop inhibitors targeting the DUBs USP7 and USP10, which are involved in regulating levels of p53 and MDM2. We were able to identify UbVs that bound USP7 or USP10 with high affinity and inhibited deubiquitination activity. We solved the crystal structure of UbV.7.2 and rationalized the molecular basis for enhanced affinity and specificity for USP7. Finally, cell death was increased significantly by UbV.7.2 expression in a colon cancer cell line that was treated with the chemotherapy drug cisplatin, demonstrating the therapeutic potential of inhibiting USP7 by this approach. (c) 2017 Elsevier Ltd. All rights reserved.