MEK1 binds directly to βarrestin1, influencing both its phosphorylation by ERK and the timing of its isoprenaline-stimulated internalization

Dong Meng, Martin J. Lynch, Elaine Huston, Michael Beyermann, Jenny Eichhorst, David R. Adams, Enno Klusmann, Miles D. Houslay, George S. Baillie

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

ßArrestin is a multifunctional signal scaffold protein. Using SPOT immobilized peptide arrays, coupled with scanning alanine substitution and mutagenesis, we show that the MAPK kinase, MEK1, interacts directly with ßarrestin1. Asp26 and Asp29 in the N-terminal domain of ßarrestin1 are critical for its binding to MEK1, whereas Arg47 and Arg49 in the N-terminal domain of MEK1 are critical for its binding to ßarrestin1. Wildtype FLAG-tagged ßarrestin1 co-immunopurifies with MEK1 in HEKB2 cells, whereas the D26A/D29A mutant does not. ERK-dependent phosphorylation at Ser412 was compromised in the D26A/D29A-ßarrestin1 mutant. A cell-permeable, 25-mer N-stearoylated ßarrestin1 peptide that encompassed the N-domain MEK1 binding site blocked ßarrestin1/ MEK1 association in HEK cells and recapitulated the altered phenotype seen with the D26A/D29A-ßarrestin1 in compromising the ERK-dependent phosphorylation of ßarrestin1. In addition, the MEK-disruptor peptide promoted the ability of ßarrestin1 to co-immunoprecipitate with endogenous c-Src and clathrin, facilitating the isoprenaline-stimulated internalization of the ß 2-adrenergic receptor. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

Original languageEnglish
Pages (from-to)11425-11435
Number of pages11
JournalJournal of Biological Chemistry
Volume284
Issue number17
DOIs
Publication statusPublished - 24 Apr 2009

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