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G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger regulatory factor via a PDZ domain-mediated interaction.

Publication ,  Journal Article
Hall, RA; Spurney, RF; Premont, RT; Rahman, N; Blitzer, JT; Pitcher, JA; Lefkowitz, RJ
Published in: J Biol Chem
August 20, 1999

The Na(+)/H(+) exchanger regulatory factor (NHERF) is constitutively phosphorylated in cells, but the site(s) of this phosphorylation and the kinase(s) responsible for it have not been identified. We show here that the primary site of constitutive NHERF phosphorylation in human embryonic kidney 293 (HEK-293) cells is Ser(289), and that the stoichiometry of phosphorylation is near 1 mol/mol. NHERF contains two PDZ domains that recognize the sequence S/T-X-L at the carboxyl terminus of target proteins, and thus we examined the possibility that kinases containing this motif might associate with and phosphorylate NHERF. Overlay experiments and co-immunoprecipitation studies revealed that NHERF binds with high affinity to a splice variant of the G protein-coupled receptor kinase 6, GRK6A, which terminates in the motif T-R-L. NHERF does not associate with GRK6B or GRK6C, alternatively spliced variants that differ from GRK6A at their extreme carboxyl termini. GRK6A phosphorylates NHERF efficiently on Ser(289) in vitro, whereas GRK6B, GRK6C, and GRK2 do not. Furthermore, the endogenous "NHERF kinase" activity in HEK-293 cell lysates is sensitive to treatments that alter the activity of GRK6A. These data suggest that GRK6A phosphorylates NHERF via a PDZ domain-mediated interaction and that GRK6A is the kinase in HEK-293 cells responsible for the constitutive phosphorylation of NHERF.

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Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

August 20, 1999

Volume

274

Issue

34

Start / End Page

24328 / 24334

Location

United States

Related Subject Headings

  • Staurosporine
  • Sodium-Hydrogen Exchangers
  • Receptor Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases
  • Phosphorylation
  • Phosphoproteins
  • Molecular Sequence Data
  • Humans
  • G-Protein-Coupled Receptor Kinases
  • Cells, Cultured
 

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Hall, R. A., Spurney, R. F., Premont, R. T., Rahman, N., Blitzer, J. T., Pitcher, J. A., & Lefkowitz, R. J. (1999). G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger regulatory factor via a PDZ domain-mediated interaction. J Biol Chem, 274(34), 24328–24334. https://doi.org/10.1074/jbc.274.34.24328
Hall, R. A., R. F. Spurney, R. T. Premont, N. Rahman, J. T. Blitzer, J. A. Pitcher, and R. J. Lefkowitz. “G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger regulatory factor via a PDZ domain-mediated interaction.J Biol Chem 274, no. 34 (August 20, 1999): 24328–34. https://doi.org/10.1074/jbc.274.34.24328.
Hall RA, Spurney RF, Premont RT, Rahman N, Blitzer JT, Pitcher JA, et al. G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger regulatory factor via a PDZ domain-mediated interaction. J Biol Chem. 1999 Aug 20;274(34):24328–34.
Hall, R. A., et al. “G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger regulatory factor via a PDZ domain-mediated interaction.J Biol Chem, vol. 274, no. 34, Aug. 1999, pp. 24328–34. Pubmed, doi:10.1074/jbc.274.34.24328.
Hall RA, Spurney RF, Premont RT, Rahman N, Blitzer JT, Pitcher JA, Lefkowitz RJ. G protein-coupled receptor kinase 6A phosphorylates the Na(+)/H(+) exchanger regulatory factor via a PDZ domain-mediated interaction. J Biol Chem. 1999 Aug 20;274(34):24328–24334.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

August 20, 1999

Volume

274

Issue

34

Start / End Page

24328 / 24334

Location

United States

Related Subject Headings

  • Staurosporine
  • Sodium-Hydrogen Exchangers
  • Receptor Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases
  • Phosphorylation
  • Phosphoproteins
  • Molecular Sequence Data
  • Humans
  • G-Protein-Coupled Receptor Kinases
  • Cells, Cultured