MEK1 binds directly to βArrestin1, influencing both its phosphorylation by ERK and the timing of its isoprenaline-stimulated internalization

Meng, D., Lynch, M. J., Huston, E., Beyermann, M., Eichhorst, J., Adams, D. R., Klusmann, E., Houslay, M. D. and Baillie, G. S. (2009) MEK1 binds directly to βArrestin1, influencing both its phosphorylation by ERK and the timing of its isoprenaline-stimulated internalization. Journal of Biological Chemistry, 284(17), pp. 11425-11435. (doi:10.1074/jbc.M806395200) (PMID:19153083) (PMCID:PMC2670148)

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Abstract

βArrestin 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 βarrestin1. Asp26 and Asp29 in the N-terminal domain of βarrestin1 are critical for its binding to MEK1, whereas Arg47 and Arg49 in the N-terminal domain of MEK1 are critical for its binding to βarrestin1. Wild-type FLAG-tagged βarrestin1 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-βarrestin1 mutant. A cell-permeable, 25-mer N-stearoylated βarrestin1 peptide that encompassed the N-domain MEK1 binding site blocked βarrestin1/MEK1 association in HEK cells and recapitulated the altered phenotype seen with the D26A/D29A-βarrestin1 in compromising the ERK-dependent phosphorylation of βarrestin1. In addition, the MEK disruptor peptide promoted the ability of βarrestin1 to co-immunoprecipitate with endogenous c-Src and clathrin, facilitating the isoprenaline-stimulated internalization of the β2-adrenergic receptor.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Baillie, Professor George and Houslay, Professor Miles and Huston, Dr Elaine
Authors: Meng, D., Lynch, M. J., Huston, E., Beyermann, M., Eichhorst, J., Adams, D. R., Klusmann, E., Houslay, M. D., and Baillie, G. S.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences > Institute of Neuroscience and Psychology
College of Medical Veterinary and Life Sciences > School of Life Sciences
Journal Name:Journal of Biological Chemistry
Journal Abbr.:J Biol Chem.
Publisher:American Society for Biochemistry and Molecular Biology, Inc.
ISSN:0021-9258
ISSN (Online):1083-351X
Published Online:19 January 2009

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
421571thera-cAMP: identification of therapeutic molecules to target compartmentalised cAMP signalling networks in human diseaseMiles HouslayEuropean Commission (EC)UNSPECIFIEDInstitute of Neuroscience and Psychology
294081PDE4 Cyclic AMP phosphodiesterasesMiles HouslayMedical Research Council (MRC)G8604010Institute of Neuroscience and Psychology
374281Protein interactions and compartmentalisation in cell signallingWalter KolchMedical Research Council (MRC)G0400053/69186Biochemistry & Cell Biology
432501Transatlantic networks of excellence in cardiovascular diseaseMiles HouslayFoundation Leducq (LEDUCQ-VIL)06 CVD 02Institute of Neuroscience and Psychology
438301Phosphodiesterase-4 isoforms - intracellular targeting, regulation and potential therapeutic targetsMiles HouslayMedical Research Council (MRC)G0600765Institute of Neuroscience and Psychology