Structure and regulation of the human Nek2 centrosomal kinase

J Biol Chem. 2007 Mar 2;282(9):6833-42. doi: 10.1074/jbc.M609721200. Epub 2006 Dec 31.

Abstract

The dimeric Ser/Thr kinase Nek2 regulates centrosome cohesion and separation through phosphorylation of structural components of the centrosome, and aberrant regulation of Nek2 activity can lead to aneuploid defects characteristic of cancer cells. Mutational analysis of autophosphorylation sites within the kinase domain identified by mass spectrometry shows a complex pattern of positive and negative regulatory effects on kinase activity that are correlated with effects on centrosomal splitting efficiency in vivo. The 2.2-A resolution x-ray structure of the Nek2 kinase domain in complex with a pyrrole-indolinone inhibitor reveals an inhibitory helical motif within the activation loop. This helix presents a steric barrier to formation of the active enzyme and generates a surface that may be exploitable in the design of specific inhibitors that selectively target the inactive state. Comparison of this "auto-inhibitory" conformation with similar arrangements in cyclin-dependent kinase 2 and epidermal growth factor receptor kinase suggests a role for dimerization-dependent allosteric regulation that combines with autophosphorylation and protein phosphatase 1c phosphatase activity to generate the precise spatial and temporal control required for Nek2 function in centrosomal maturation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Regulation*
  • Binding Sites
  • Centrosome / enzymology*
  • Crystallography, X-Ray
  • DNA Mutational Analysis
  • Dimerization
  • Humans
  • Mass Spectrometry
  • NIMA-Related Kinases
  • Phosphorylation
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein Serine-Threonine Kinases / chemistry*

Substances

  • NEK2 protein, human
  • NIMA-Related Kinases
  • Protein Serine-Threonine Kinases

Associated data

  • PDB/2JAV