USP7-specific inhibitors target and modify the enzyme’s active site via distinct chemical mechanisms.
Open accessAbstract
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.
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Mutation of C223 to alanine prevents inhibitor binding to USP7, revealing specificity for the catalytic cysteine.
“Here, mutation of C223 to an alanine prevents inhibitor binding to USP7, revealing specificity for the catalytic cysteine (Pozhidaeva et al., 2017).”
From USP7 Is a Master Regulator of Genome Stability (2020) · cites this paper as Pozhidaeva et al., 2017
P22077 and P50429, derivatives of 5091, both irreversibly modify C223.
“P22077 and P50429, also derivatives of 5091, both irreversibly modify C223.”
From USP7 Is a Master Regulator of Genome Stability (2020) · cites this paper as Pozhidaeva et al., 2017
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USP7: Structure, substrate specificity, and inhibition
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Self-citationUSP7 Is a Master Regulator of Genome Stability
Valles, G. J., Bezsonova, I., Woodgate, R., & Ashton, N. W. (2020) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2020.00717 · Cited by 114 · see citing works
“The C-terminal tail of ubiquitin conjugated to a lysine residue of the substrate enters this cleft, allowing for hydrolysis of the isopeptide bond between ubiquitin and the substrate (Hu et al., 2001; Pozhidaeva et al., 2017).”
BackgroundSelf-citationKinetic analysis of multistep USP7 mechanism shows critical role for target protein in activity.
Kim, R. Q., Geurink, P. P., Mulder, M. P. C., Fish, A., Ekkebus, R., El Oualid, F., van Dijk, W. J., van Dalen, D., Ovaa, H., van Ingen, H., et al. (2019) · Nature Communications · doi:10.1038/s41467-018-08231-5 · Cited by 41 · see citing works
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Disulfiram and 6-Thioguanine synergistically inhibit the enzymatic activities of USP2 and USP21
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- Authors
- Pozhidaeva, A., Valles, G., Wang, F., Wu, J., Sterner, D. E., Nguyen, P., Weinstock, J., Kumar, K. G. S., Kanyo, J., Wright, D., & Bezsonova, I.
- Affiliations
- UConn Health (US)
- Progenra (United States) (US)
- Yale University (US)
- University of Connecticut (US)
- Corresponding author
- Bezsonova, Irina
- ORCID
- Venue
- Cell Chemical Biology
- Publisher
- Elsevier BV
- Published
- 2017-12
- Volume
- 24
- Issue
- 12
- Pages
- 1501-1512.e5
- ISSN
- 2451-9448 (ISSN-L)
- 2451-9456
- OpenAlex
- W2765710196
- Cited by
- 107
- References
- 58
- Citations by year
- 2026: 7 · 2025: 16 · 2024: 8 · 2023: 9 · 2022: 13 · 2021: 16 · 2020: 18 · 2019: 10 · 2018: 9 · 2017: 1
- FWCI
- 4.25
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Cell chemical biology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- NSF (1616184)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Protease and Inhibitor Mechanisms
- Keywords
- USP7, ubiquitin binding, conformational switching, enzyme activation, anticancer drug development, oncogenic signaling
- MeSH terms
- Ubiquitin-Specific Peptidase 7, Humans, Models, Molecular, Protease Inhibitors, Structure-Activity Relationship, Substrate Specificity, Thiophenes, Molecular Structure, Catalytic Domain, Ubiquitin
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