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

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Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction. / Cardoso, Rosa; Love, Robert; Nilsson, Carol L; Bergqvist, Simon; Nowlin, Dawn; Yan, Jiangli; Liu, Kevin K-C; Zhu, Jing; Chen, Ping; Deng, Ya-Li; Dyson, H Jane; Greig, Michael J; Brooun, Alexei.

I: Protein Science, Vol. 21, Nr. 12, 12.2012, s. 1885-96.

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

Harvard

Cardoso, R, Love, R, Nilsson, CL, Bergqvist, S, Nowlin, D, Yan, J, Liu, KK-C, Zhu, J, Chen, P, Deng, Y-L, Dyson, HJ, Greig, MJ & Brooun, A 2012, 'Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction', Protein Science, vol. 21, nr. 12, s. 1885-96. https://doi.org/10.1002/pro.2172

APA

Cardoso, R., Love, R., Nilsson, C. L., Bergqvist, S., Nowlin, D., Yan, J., Liu, K. K-C., Zhu, J., Chen, P., Deng, Y-L., Dyson, H. J., Greig, M. J., & Brooun, A. (2012). Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction. Protein Science, 21(12), 1885-96. https://doi.org/10.1002/pro.2172

CBE

Cardoso R, Love R, Nilsson CL, Bergqvist S, Nowlin D, Yan J, Liu KK-C, Zhu J, Chen P, Deng Y-L, Dyson HJ, Greig MJ, Brooun A. 2012. Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction. Protein Science. 21(12):1885-96. https://doi.org/10.1002/pro.2172

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Author

Cardoso, Rosa ; Love, Robert ; Nilsson, Carol L ; Bergqvist, Simon ; Nowlin, Dawn ; Yan, Jiangli ; Liu, Kevin K-C ; Zhu, Jing ; Chen, Ping ; Deng, Ya-Li ; Dyson, H Jane ; Greig, Michael J ; Brooun, Alexei. / Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction. I: Protein Science. 2012 ; Vol. 21, Nr. 12. s. 1885-96.

RIS

TY - JOUR

T1 - Identification of Cys255 in HIF-1α as a novel site for development of covalent inhibitors of HIF-1α/ARNT PasB domain protein-protein interaction

AU - Cardoso, Rosa

AU - Love, Robert

AU - Nilsson, Carol L

AU - Bergqvist, Simon

AU - Nowlin, Dawn

AU - Yan, Jiangli

AU - Liu, Kevin K-C

AU - Zhu, Jing

AU - Chen, Ping

AU - Deng, Ya-Li

AU - Dyson, H Jane

AU - Greig, Michael J

AU - Brooun, Alexei

N1 - Copyright © 2012 The Protein Society.

PY - 2012/12

Y1 - 2012/12

N2 - 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.

AB - 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.

KW - Aryl Hydrocarbon Receptor Nuclear Translocator

KW - Cysteine

KW - Humans

KW - Hypoxia-Inducible Factor 1, alpha Subunit

KW - Models, Molecular

KW - Protein Binding

KW - Protein Conformation

KW - Protein Interaction Domains and Motifs

KW - Protein Interaction Maps

KW - Protein Multimerization

KW - Small Molecule Libraries

KW - Journal Article

U2 - 10.1002/pro.2172

DO - 10.1002/pro.2172

M3 - Article

C2 - 23033253

VL - 21

SP - 1885

EP - 1896

JO - Protein Science

JF - Protein Science

SN - 1469-896X

IS - 12

ER -