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PDBsum entry 3hmp
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References listed in PDB file
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Key reference
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Title
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Biophysical and X-Ray crystallographic analysis of mps1 kinase inhibitor complexes.
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Authors
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M.L.Chu,
Z.Lang,
L.M.Chavas,
J.Neres,
O.S.Fedorova,
L.Tabernero,
M.Cherry,
D.H.Williams,
K.T.Douglas,
P.A.Eyers.
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Ref.
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Biochemistry, 2010,
49,
1689-1701.
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PubMed id
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Abstract
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The dual-specificity protein kinase monopolar spindle 1 (Mps1) is a central
component of the mitotic spindle assembly checkpoint (SAC), a sensing mechanism
that prevents anaphase until all chromosomes are bioriented on the metaphase
plate. Partial depletion of Mps1 protein levels sensitizes transformed, but not
untransformed, human cells to therapeutic doses of the anticancer agent Taxol,
making it an attractive novel therapeutic cancer target. We have previously
determined the X-ray structure of the catalytic domain of human Mps1 in complex
with the anthrapyrazolone kinase inhibitor SP600125. In order to validate
distinct inhibitors that target this enzyme and improve our understanding of
nucleotide binding site architecture, we now report a biophysical and structural
evaluation of the Mps1 catalytic domain in the presence of ATP and the aspecific
model kinase inhibitor staurosporine. Collective in silico, enzymatic, and
fluorescent screens also identified several new lead quinazoline Mps1
inhibitors, including a low-affinity compound termed Compound 4 (Cpd 4), whose
interaction with the Mps1 kinase domain was further characterized by X-ray
crystallography. A novel biophysical analysis demonstrated that the intrinsic
fluorescence of SP600125 changed markedly upon Mps1 binding, allowing
spectrophotometric displacement analysis and determination of dissociation
constants for ATP-competitive Mps1 inhibitors. By illuminating the structure of
the Mps1 ATP-binding site our results provide novel biophysical insights into
Mps1-ligand interactions that will be useful for the development of specific
Mps1 inhibitors, including those employing a therapeutically validated
quinazoline template.
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