PDBsum entry 4h6j

Transcription

PDB id: 4h6j

Contents

Protein chains

106 a.a.

111 a.a.

Waters ×154

PDB id:

4h6j

Name:

Transcription

Title:

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.

Structure:

Hypoxia inducible factor 1-alpha. Chain: a. Fragment: pas-b domain, unp residues 238-348. Engineered: yes. Mutation: yes. Aryl hydrocarbon nuclear translocator. Chain: b. Fragment: pas-b domain, unp residues 357-470. Engineered: yes.

Source:

Homo sapiens. Organism_taxid: 9606. Gene: hif-1 alpha. Expressed in: escherichia coli. Expression_system_taxid: 511693. Gene: arnt.

Resolution:

1.52Å R-factor: 0.216 R-free: 0.248

Authors:

R.Cardoso,R.A.Love,C.Nilsson,S.Bergqvist,D.Nowlin,J.Yan,K.Liu,J.Zhu, P.Chen,Y.-L.Deng,H.J.Dyson,M.J.Greig,A.Brooun

Key ref:

R.Cardoso et al. (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 Sci, 21, 1885-1896. PubMed id: 23033253

Date:

19-Sep-12 Release date: 05-Dec-12

Protein chain

Q16665  (HIF1A_HUMAN) -  Hypoxia-inducible factor 1-alpha from Homo sapiens

Seq: 826 a.a.

Struc: 106 a.a.*

Protein chain

P27540  (ARNT_HUMAN) -  Aryl hydrocarbon receptor nuclear translocator from Homo sapiens

Seq: 789 a.a.

Struc: 111 a.a.*

* PDB and UniProt seqs differ at 3 residue positions (black crosses)

Enzyme reactions

• Enzyme class: Chains A, B: E.C.?

Protein Sci 21:1885-1896 (2012)

PubMed id: 23033253

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

R.Cardoso, R.Love, C.L.Nilsson, S.Bergqvist, D.Nowlin, J.Yan, K.K.Liu, J.Zhu, P.Chen, Y.L.Deng, H.J.Dyson, M.J.Greig, A.Brooun.

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.