Vimentin Binding to Phosphorylated Erk Sterically Hinders Enzymatic Dephosphorylation of the Kinase

Eran Perlson, Izhak Michaelevski, Noga Kowalsman, Keren Ben-Yaakov, Maya Shaked, Rony Seger, Miriam Eisenstein, Mike Fainzilber*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Cleavage fragments of de novo synthesized vimentin were recently reported to interact with phosphorylated Erk1 and Erk2 MAP kinases (pErk) in injured sciatic nerve, thus linking pErk to a signaling complex retrogradely transported on importins and dynein. Here we clarify the structural basis for this interaction, which explains how pErk is protected from dephosphorylation while bound to vimentin. Pull-down and ELISA experiments revealed robust calcium-dependent binding of pErk to the second coiled-coil domain of vimentin, with observed affinities of binding increasing from 180 nM at 0.1 μM calcium to 15 nM at 10 μM calcium. In contrast there was little or no binding of non-phosphorylated Erk to vimentin under these conditions. Geometric and electrostatic complementarity docking generated a number of solutions wherein vimentin binding to pErk occludes the lip containing the phosphorylated residues in the kinase. Binding competition experiments with Erk peptides confirmed a solution in which vimentin covers the phosphorylation lip in pErk, interacting with residues above and below the lip. The same peptides inhibited pErk binding to the dynein complex in sciatic nerve axoplasm, and interfered with protection from phosphatases by vimentin. Thus, a soluble intermediate filament fragment interacts with a signaling kinase and protects it from dephosphorylation by calcium-dependent steric hindrance.

Original languageEnglish
Pages (from-to)938-944
Number of pages7
JournalJournal of Molecular Biology
Volume364
Issue number5
DOIs
StatePublished - 15 Dec 2006
Externally publishedYes

Keywords

  • Erk
  • docking
  • intermediate filament
  • phosphorylation
  • vimentin

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