Regulation of g protein-coupled receptor kinases by calmodulin and localization of the calmodulin binding domain

A. N. Pronin, D. K. Satpaev, Vladlen Z Slepak, J. L. Benovic

Research output: Contribution to journalArticle

Abstract

G protein-coupled receptor kinases (GRKs) specifically phosphorylate and regulate the activated form of multiple G protein-coupled receptors. Recent studies have revealed that GRKs are also subject to regulation. For example, GRK2 and GRK5 can be phosphorylated and either activated or inhibited, respectively, by protein kinase C. Here we demonstrate that calmodulin is a potent inhibitor of GRK activity with a selectivity for GRK5 (ICs0 50 nM) > GRK6 >> GRK2 (ICso 2000 nM) >> GRK1. Calmodulin inhibition of GRK5 is mediated via a reduced kinase binding to both receptor and phospholipid. Interestingly, calmodulin activates autophosphorylation of GRK5 at sites distinct from the two major autophosphorylation sites on GRK5. Moreover. calmodulin-stimulated autophosphorylation of GRK5 directly inhibits GRK5 interaction with the receptor even in the absence of calmodulin. Using gluthatione-S-transferase-GRK5 fusion proteins to either inhibit calmodulinstimulated autophosphorylation or bind directly to calmodulin, we determined that an amino terminal domain of GRK5 (amino acids 20-39) is sufficient for calmodulin binding. This domain is abundant in basic and hydrophobic residues, characteristics typical of calmodulin binding sites, and is highly conserved in GRK4, GRK5, and GRK6. These studies suggest that calmodulin may serve a general role in mediating calcium-dependent regulation of GRK activity. Supported by grants from NIH, Dept. of the Navy, and the American Heart Association.

Original languageEnglish
JournalFASEB Journal
Volume11
Issue number9
StatePublished - Dec 1 1997
Externally publishedYes

Fingerprint

calmodulin
Calmodulin
phosphotransferases (kinases)
Phosphotransferases
G-Protein-Coupled Receptor Kinases
receptors
protein phosphorylation
Proteins
proteins
Organized Financing
protein kinase C
Transferases
G-Protein-Coupled Receptors
transferases
Protein Kinase C
binding sites
Phospholipids
phospholipids
Fusion reactions
Binding Sites

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Regulation of g protein-coupled receptor kinases by calmodulin and localization of the calmodulin binding domain. / Pronin, A. N.; Satpaev, D. K.; Slepak, Vladlen Z; Benovic, J. L.

In: FASEB Journal, Vol. 11, No. 9, 01.12.1997.

Research output: Contribution to journalArticle

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