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Cell Chem Biol. 2017 Dec 21;24(12):1501-1512.e5. doi: 10.1016/j.chembiol.2017.09.004. Epub 2017 Oct 19.

USP7-Specific Inhibitors Target and Modify the Enzyme's Active Site via Distinct Chemical Mechanisms.

Author information

1
Department of Molecular Biology and Biophysics, UConn Health, Farmington, CT 06030, USA.
2
Progenra Inc., Malvern, PA 19355, USA.
3
Keck Foundation Biotechnology Resource Laboratory, Yale University, New Haven, CT 06511, USA.
4
Department of Pharmaceutical Sciences, University of Connecticut, Storrs, CT 06268, USA.
5
Department of Molecular Biology and Biophysics, UConn Health, Farmington, CT 06030, USA. Electronic address: bezsonova@uchc.edu.

Abstract

USP7 is a deubiquitinating enzyme that plays a pivotal role in multiple oncogenic pathways and therefore is a desirable target for new anti-cancer therapies. However, the lack of structural information about the USP7-inhibitor interactions has been a critical gap in the development of potent inhibitors. USP7 is unique among USPs in that its active site is catalytically incompetent, and is postulated to rearrange into a productive conformation only upon binding to ubiquitin. Surprisingly, we found that ubiquitin alone does not induce an active conformation in solution. Using a combination of nuclear magnetic resonance, mass spectrometry, computational modeling, and cell-based assays, we found that DUB inhibitors P22077 and P50429 covalently modify the catalytic cysteine of USP7 and induce a conformational switch in the enzyme associated with active site rearrangement. This work represents the first experimental insights into USP7 activation and inhibition and provides a structural basis for rational development of potent anti-cancer therapeutics.

KEYWORDS:

DUB inhibition; DUBs; NMR; USP7; covalent inhibitors; cysteine peptidase; mass spectrometry; ubiquitin binding

PMID:
29056420
DOI:
10.1016/j.chembiol.2017.09.004
[Indexed for MEDLINE]
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