4H6J

Identification of Cys 255 in HIF-1 as a novel site for development of covalent inhibitors of HIF-1 /ARNT PasB domain protein-protein interaction.

Summary for 4H6J

• Entry DOI: 10.2210/pdb4h6j/pdb
• Related: 3F1P
• Descriptor: HYPOXIA INDUCIBLE FACTOR 1-ALPHA, ARYL HYDROCARBON NUCLEAR TRANSLOCATOR (3 entities in total)
• Functional Keywords: pas domain, heterodimer, hypoxia inducible factor, transcription factor, arnt, transcription
• Biological source: Homo sapiens; More
• Cellular location: Cytoplasm : Q16665; Nucleus: P27540
• Total number of polymer chains: 2
• Total formula weight: 26692.05

Authors

Cardoso, R.,Love, R.A.,Nilsson, C.,Bergqvist, S.,Nowlin, D.,Yan, J.,Liu, K.,Zhu, J.,Chen, P.,Deng, Y.-L.,Dyson, H.J.,Greig, M.J.,Brooun, A. (deposition date: 2012-09-19, release date: 2012-12-05, Last modification date: 2024-02-28)

Primary citation

Cardoso, R.,Love, R.,Nilsson, C.L.,Bergqvist, S.,Nowlin, D.,Yan, J.,Liu, K.K.,Zhu, J.,Chen, P.,Deng, Y.L.,Dyson, H.J.,Greig, M.J.,Brooun, A.
Identification of Cys255 in HIF-1 alpha as a novel site for development of covalent inhibitors of HIF-1 alpha /ARNT PasB domain protein-protein interaction.
Protein Sci., 21:1885-1896, 2012

PubMed 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.

PubMed: 23033253
DOI: 10.1002/pro.2172

Experimental method

X-RAY DIFFRACTION (1.52 Å)