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

Authors

Andrea Piserchio, CUNY City College
Venkatesh Ramakrishan, CUNY City College
Hsin Wang, CUNY City College
Tamer S. Kaoud, University of Texas, Austin
Boris Arshava, CUNY College of Staten Island
Kaushik Dutta, The New York Structural Biology Center
Kevin N. Dalby, University of Texas, Austin
Ranajeet Ghose, CUNY City College

Document Type

Article

Publication Date

June 2015

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.

Comments

This work was originally published in Scientific Reports, available at doi:10.1038/srep11127.