Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction

Research output: Contribution to journalArticle


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",
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 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.",
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",
author = "Rosa Cardoso and Robert Love and Nilsson, {Carol L} and Simon Bergqvist and Dawn Nowlin and Jiangli Yan and Liu, {Kevin K-C} and Jing Zhu and Ping Chen and Ya-Li Deng and Dyson, {H Jane} and Greig, {Michael J} and Alexei Brooun",
note = "Copyright {\textcopyright} 2012 The Protein Society.",
year = "2012",
month = dec,
doi = "10.1002/pro.2172",
language = "English",
volume = "21",
pages = "1885--96",
journal = "Protein Science",
issn = "1469-896X",
publisher = "The Protein Society",
number = "12",