Structural and Dynamic Features of F-recruitment Site Driven Substrate Phosphorylation by ERK2

Sci Rep. 2015 Jun 8;5:11127. doi: 10.1038/srep11127.

Abstract

The F-recruitment site (FRS) of active ERK2 binds F-site (Phe-x-Phe-Pro) sequences found downstream of the Ser/Thr phospho-acceptor on cellular substrates. Here we apply NMR methods to analyze the interaction between active ERK2 (ppERK2), and a 13-residue F-site-bearing peptide substrate derived from its cellular target, the transcription factor Elk-1. Our results provide detailed insight into previously elusive structural and dynamic features of FRS/F-site interactions and FRS-driven substrate phosphorylation. We show that substrate F-site engagement significantly quenches slow dynamics involving the ppERK2 activation-loop and the FRS. We also demonstrate that the F-site phenylalanines make critical contacts with ppERK2, in contrast to the proline whose cis-trans isomerization has no significant effect on F-site recognition by the kinase FRS. Our results support a mechanism where phosphorylation of the disordered N-terminal phospho-acceptor is facilitated by its increased productive encounters with the ppERK2 active site due to docking of the proximal F-site at the kinase FRS.

Publication types

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

MeSH terms

  • Binding Sites / physiology
  • Catalytic Domain / physiology*
  • DNA / metabolism
  • Humans
  • Mitogen-Activated Protein Kinase 1 / metabolism*
  • Models, Molecular
  • Phosphorylation
  • Protein Conformation
  • ets-Domain Protein Elk-1 / metabolism*

Substances

  • ELK1 protein, human
  • ets-Domain Protein Elk-1
  • DNA
  • MAPK1 protein, human
  • Mitogen-Activated Protein Kinase 1