PDBsum entry 2owb

Transferase

PDB id: 2owb

Contents

Protein chain

294 a.a. *

Ligands

ACT ×2

626

Metals

_ZN

Waters ×256

* Residue conservation analysis

PDB id:

2owb

Name:

Transferase

Title:

Structure of the catalytic domain of human polo-like kinase 1

Structure:

Serine/threonine-protein kinase plk1. Chain: a. Fragment: kinase domain (residues 13-345). Synonym: polo-like kinase 1, plk-1, serine/threonine-protein kinase 13, stpk13. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: plk1, plk. Expressed in: spodoptera frugiperda. Expression_system_taxid: 7108.

Resolution:

2.10Å R-factor: 0.205 R-free: 0.243

Authors:

Y.-H.Ding,M.Kothe,D.Kohls,S.Low

Key ref:

M.Kothe et al. (2007). Structure of the catalytic domain of human polo-like kinase 1. Biochemistry, 46, 5960-5971. PubMed id: 17461553

Date:

15-Feb-07 Release date: 24-Apr-07

Protein chain

P53350  (PLK1_HUMAN) -  Serine/threonine-protein kinase PLK1 from Homo sapiens

Seq: 603 a.a.

Struc: 294 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.2.7.11.21 - polo kinase.
• Reaction:
  1. L-seryl-[protein] + ATP = O-phospho-L-seryl-[protein] + ADP + H⁺
  2. L-threonyl-[protein] + ATP = O-phospho-L-threonyl-[protein] + ADP + H⁺

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

Biochemistry 46:5960-5971 (2007)

PubMed id: 17461553

Structure of the catalytic domain of human polo-like kinase 1.

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.

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.