skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10

Abstract

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 inmore » a colon cancer cell line that was treated with the chemotherapy drug cisplatin, demonstrating the therapeutic potential of inhibiting USP7 by this approach« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
FOREIGN
OSTI Identifier:
1410112
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Molecular Biology; Journal Volume: 429; Journal Issue: 22
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; USP7; USP10; p53; inhibitor design

Citation Formats

Zhang, Wei, Sartori, Maria A., Makhnevych, Taras, Federowicz, Kelly E., Dong, Xiaohui, Liu, Li, Nim, Satra, Dong, Aiping, Yang, Jingsong, Li, Yanjun, Haddad, Dania, Ernst, Andreas, Heerding, Dirk, Tong, Yufeng, Moffat, Jason, and Sidhu, Sachdev S. Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10. United States: N. p., 2017. Web. doi:10.1016/j.jmb.2017.05.025.
Zhang, Wei, Sartori, Maria A., Makhnevych, Taras, Federowicz, Kelly E., Dong, Xiaohui, Liu, Li, Nim, Satra, Dong, Aiping, Yang, Jingsong, Li, Yanjun, Haddad, Dania, Ernst, Andreas, Heerding, Dirk, Tong, Yufeng, Moffat, Jason, & Sidhu, Sachdev S. Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10. United States. doi:10.1016/j.jmb.2017.05.025.
Zhang, Wei, Sartori, Maria A., Makhnevych, Taras, Federowicz, Kelly E., Dong, Xiaohui, Liu, Li, Nim, Satra, Dong, Aiping, Yang, Jingsong, Li, Yanjun, Haddad, Dania, Ernst, Andreas, Heerding, Dirk, Tong, Yufeng, Moffat, Jason, and Sidhu, Sachdev S. Wed . "Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10". United States. doi:10.1016/j.jmb.2017.05.025.
@article{osti_1410112,
title = {Generation and Validation of Intracellular Ubiquitin Variant Inhibitors for USP7 and USP10},
author = {Zhang, Wei and Sartori, Maria A. and Makhnevych, Taras and Federowicz, Kelly E. and Dong, Xiaohui and Liu, Li and Nim, Satra and Dong, Aiping and Yang, Jingsong and Li, Yanjun and Haddad, Dania and Ernst, Andreas and Heerding, Dirk and Tong, Yufeng and Moffat, Jason and Sidhu, Sachdev S.},
abstractNote = {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},
doi = {10.1016/j.jmb.2017.05.025},
journal = {Journal of Molecular Biology},
number = 22,
volume = 429,
place = {United States},
year = {Wed Nov 01 00:00:00 EDT 2017},
month = {Wed Nov 01 00:00:00 EDT 2017}
}