Identification of Cys255 in HIF‐1α as a novel site for development of covalent inhibitors of HIF‐1α/ARNT PasB domain protein–protein interaction
Open accessAbstract
The heterodimer HIF‐1α (hypoxia inducible factor)/HIF‐β (also known as ARNT‐aryl hydrocarbon nuclear translocator) is a key mediator of cellular response to hypoxia. The interaction between these monomer units can be modified by the action of small molecules in the binding interface between their C‐terminal heterodimerization (PasB) domains. Taking advantage of the presence of several cysteine residues located in the allosteric cavity of HIF‐1α PasB domain, we applied a cysteine‐based reactomics “hotspot identification” strategy to locate regions of HIF‐1α PasB domain critical for its interaction with ARNT. COMPOUND 5 was identified using a mass spectrometry‐based primary screening strategy and was shown to react specifically with Cys255 of the HIF‐1α PasB domain. Biophysical characterization of the interaction between PasB domains of HIF‐1α and ARNT revealed that covalent binding of COMPOUND 5 to Cys255 reduced binding affinity between HIF‐1α and ARNT PasB domains approximately 10‐fold. Detailed NMR structural analysis of HIF‐1α‐PasB‐COMPOUND 5 conjugate showed significant local conformation changes in the HIF‐1α associated with key residues involved in the HIF‐1α/ARNT PasB domain interaction as revealed by the crystal structure of the HIF‐1α/ARNT PasB heterodimer. Our screening strategy could be applied to other targets to identify pockets surrounding reactive cysteines suitable for development of small molecule modulators of protein function.
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In the specific field of oncology, one covalent PPI inhibitor has been approved by the FDA, while others have entered clinical trials.
“In the specific field of oncology, one covalent PPI inhibitor has been approved by the FDA, while others have entered clinical trials [84, 89, 91–93].”
From The design and development of covalent protein-protein interaction inhibitors for cancer treatment (2020) · cites this paper as [84]
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- Authors
- Cardoso, R., Love, R., Nilsson, C. L., Bergqvist, S., Nowlin, D., Yan, J., Liu, K. K. ‐., Zhu, J., Chen, P., Deng, Y., Dyson, H. J., Greig, M. J., & Brooun, A.
- Affiliations
- Pfizer (United States) (US)
- Scripps Research Institute (US)
- Corresponding author
- Brooun, Alexei
- Venue
- Protein Science
- Publisher
- Wiley
- Published
- 2012-11-09
- Volume
- 21
- Issue
- 12
- Pages
- 1885-1896
- ISSN
- 0961-8368 (ISSN-L)
- 1469-896X
- PMC
- PMC3575918
- OpenAlex
- W2165699712
- Cited by
- 79
- References
- 43
- Citations by year
- 2026: 2 · 2025: 9 · 2024: 7 · 2023: 4 · 2022: 8 · 2021: 8 · 2020: 7 · 2019: 4 · 2018: 5 · 2017: 7 · 2016: 8 · 2015: 7 · 2014: 2 · 2013: 1
- FWCI
- 1.06
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Protein Science (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Lund University Publications (Lund University) (submittedVersion)
- Funders
- National Institute of General Medical Sciences (P01GM071862)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Physical Sciences › Chemistry › Organic Chemistry › Synthesis and biological activity
- Keywords
- HIF-1α, ARNT, protein-protein interaction, covalent inhibitors, hotspot detection, hypoxia, small molecule modulators, protein conformational change
- MeSH terms
- Cysteine, Humans, Models, Molecular, Protein Binding, Protein Conformation, Aryl Hydrocarbon Receptor Nuclear Translocator, Hypoxia-Inducible Factor 1, alpha Subunit, Protein Interaction Domains and Motifs, Small Molecule Libraries, Protein Multimerization, Protein Interaction Maps
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