title: Structure of the catalytic domain of human polo-like kinase 1.
abstract: 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.
authors.affiliation: Pfizer Global Research and Development, Research Technology Center, 620 Memorial Drive, Cambridge, Massachusetts 02139, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Kothe
authors.forename: Darcy
authors.initials: D
authors.lastname: Kohls
authors.forename: Simon
authors.initials: S
authors.lastname: Low
authors.forename: Rocco
authors.initials: R
authors.lastname: Coli
authors.forename: Alan C
authors.initials: AC
authors.lastname: Cheng
authors.forename: Suzanne L
authors.initials: SL
authors.lastname: Jacques
authors.forename: Theresa L
authors.initials: TL
authors.lastname: Johnson
authors.forename: Cristina
authors.initials: C
authors.lastname: Lewis
authors.forename: Christine
authors.initials: C
authors.lastname: Loh
authors.forename: Jim
authors.initials: J
authors.lastname: Nonomiya
authors.forename: Alissa L
authors.initials: AL
authors.lastname: Sheils
authors.forename: Kimberly A
authors.initials: KA
authors.lastname: Verdries
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Wynn
authors.forename: Cyrille
authors.initials: C
authors.lastname: Kuhn
authors.forename: Yuan-Hua
authors.initials: YH
authors.lastname: Ding
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018360:Crystallography, X-Ray; D004789:Enzyme Activation / Q000235:genetics; D004795:Enzyme Stability / Q000235:genetics; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011487:Protein Conformation; D017510:Protein Folding; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000097869:Polo-Like Kinase 1
chemical_list: D018797:Cell Cycle Proteins; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D000097869:Polo-Like Kinase 1