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PDBsum entry 4h6j
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Transcription
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PDB id
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4h6j
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References listed in PDB file
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Key reference
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Title
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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.
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Authors
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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.
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Ref.
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Protein Sci, 2012,
21,
1885-1896.
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PubMed id
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Abstract
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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.
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