Solution NMR insights into docking interactions involving inactive ERK2

Biochemistry. 2011 May 10;50(18):3660-72. doi: 10.1021/bi2000559. Epub 2011 Apr 19.


The mitogen-activated protein (MAP) kinase ERK2 contains recruitment sites that engage canonical and noncanonical motifs found in a variety of upstream kinases, regulating phosphatases and downstream targets. Interactions involving two of these sites, the D-recruitment site (DRS) and the F-recruitment site (FRS), have been shown to play a key role in signal transduction by ERK/MAP kinases. The dynamic nature of these recruitment events makes NMR uniquely suited to provide significant insight into these interactions. While NMR studies of kinases in general have been greatly hindered by their large size and complex dynamic behavior leading to the suboptimal performance of standard methodologies, we have overcome these difficulties for inactive full-length ERK2 and obtained an acceptable level of backbone resonance assignments. This allowed a detailed investigation of the structural perturbations that accompany interactions involving both canonical and noncanonical recruitment events. No crystallographic information exists for the latter. We found that the chemical shift perturbations in inactive ERK2, indicative of structural changes in the presence of canonical and noncanonical motifs, are not restricted to the recruitment sites but also involve the linker that connects the N- and C-lobes and, in most cases, a gatekeeper residue that is thought to exert allosteric control over catalytic activity. We also found that, even though the canonical motifs interact with the DRS utilizing both charge-charge and hydrophobic interactions, the noncanonical interactions primarily involve the latter. These results demonstrate the feasibility of solution NMR techniques for a comprehensive analysis of docking interactions in a full-length ERK/MAP kinase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Site
  • Amino Acid Motifs
  • Animals
  • Binding Sites
  • Crystallography, X-Ray / methods
  • Escherichia coli / metabolism
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Humans
  • MAP Kinase Signaling System
  • Magnetic Resonance Spectroscopy / methods*
  • Mitogen-Activated Protein Kinase 1 / chemistry*
  • Models, Statistical
  • Molecular Conformation
  • Peptides / chemistry
  • Protein Binding
  • Protein Structure, Tertiary
  • Rats


  • Peptides
  • Extracellular Signal-Regulated MAP Kinases
  • MAPK1 protein, human
  • Mitogen-Activated Protein Kinase 1