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PDBsum entry 2owb
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References listed in PDB file
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Key reference
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Title
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Structure of the catalytic domain of human polo-Like kinase 1.
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Authors
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M.Kothe,
D.Kohls,
S.Low,
R.Coli,
A.C.Cheng,
S.L.Jacques,
T.L.Johnson,
C.Lewis,
C.Loh,
J.Nonomiya,
A.L.Sheils,
K.A.Verdries,
T.A.Wynn,
C.Kuhn,
Y.H.Ding.
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Ref.
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Biochemistry, 2007,
46,
5960-5971.
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PubMed id
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Abstract
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Polo-like kinase 1 (Plk1) is an attractive target for the development of
anticancer agents due to its importance in regulating cell-cycle progression.
Overexpression of Plk1 has been detected in a variety of cancers, and expression
levels often correlate with poor prognosis. Despite high interest in
Plk1-targeted therapeutics, there is currently no structure publicly available
to guide structure-based drug design of specific inhibitors. We determined the
crystal structures of the T210V mutant of the kinase domain of human Plk1
complexed with the nonhydrolyzable ATP analogue adenylylimidodiphosphate
(AMPPNP) or the pyrrolo-pyrazole inhibitor PHA-680626 at 2.4 and 2.1 A
resolution, respectively. Plk1 adopts the typical kinase domain fold and
crystallized in a conformation resembling the active state of other kinases.
Comparison of the kinetic parameters determined for the (unphosphorylated)
wild-type enzyme, as well as the T210V and T210D mutants, shows that the
mutations primarily affect the kcat of the reaction, with little change in the
apparent Km for the protein or nucleotide substrates (kcat = 0.0094, 0.0376, and
0.0049 s-1 and Km(ATP) = 3.2, 4.0, and 3.0 microM for WT, T210D, and T210V,
respectively). The structure highlights features of the active site that can be
exploited to obtain Plk1-specific inhibitors with selectivity over other kinases
and Plk isoforms. These include the presence of a phenylalanine at the bottom of
the ATP pocket, combined with a cysteine (as opposed to the more commonly found
leucine) in the roof of the binding site, a pocket created by Leu132 in the
hinge region, and a cluster of positively charged residues in the
solvent-exposed area outside of the adenine pocket adjacent to the hinge region.
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