Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction
(2012) In Protein Science 21(12). p.96-1885- Abstract
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... (More)
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.
(Less)
- author
- publishing date
- 2012-12
- type
- Contribution to journal
- publication status
- published
- keywords
- Aryl Hydrocarbon Receptor Nuclear Translocator, Cysteine, Humans, Hypoxia-Inducible Factor 1, alpha Subunit, Models, Molecular, Protein Binding, Protein Conformation, Protein Interaction Domains and Motifs, Protein Interaction Maps, Protein Multimerization, Small Molecule Libraries, Journal Article
- in
- Protein Science
- volume
- 21
- issue
- 12
- pages
- 12 pages
- publisher
- The Protein Society
- external identifiers
-
- pmid:23033253
- scopus:84870354792
- ISSN
- 1469-896X
- DOI
- 10.1002/pro.2172
- language
- English
- LU publication?
- no
- id
- 9edbdc34-2697-4c63-b090-a127497a9b73
- date added to LUP
- 2017-05-16 10:28:28
- date last changed
- 2025-05-12 22:29:55
@article{9edbdc34-2697-4c63-b090-a127497a9b73,
abstract = {{<p>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.</p>}},
author = {{Cardoso, Rosa and Love, Robert and Nilsson, Carol L and Bergqvist, Simon and Nowlin, Dawn and Yan, Jiangli and Liu, Kevin K-C and Zhu, Jing and Chen, Ping and Deng, Ya-Li and Dyson, H Jane and Greig, Michael J and Brooun, Alexei}},
issn = {{1469-896X}},
keywords = {{Aryl Hydrocarbon Receptor Nuclear Translocator; Cysteine; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Models, Molecular; Protein Binding; Protein Conformation; Protein Interaction Domains and Motifs; Protein Interaction Maps; Protein Multimerization; Small Molecule Libraries; Journal Article}},
language = {{eng}},
number = {{12}},
pages = {{96--1885}},
publisher = {{The Protein Society}},
series = {{Protein Science}},
title = {{Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction}},
url = {{http://dx.doi.org/10.1002/pro.2172}},
doi = {{10.1002/pro.2172}},
volume = {{21}},
year = {{2012}},
}