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Isoprenylation of a protein kinase. Requirement of farnesylation/alpha-carboxyl methylation for full enzymatic activity of rhodopsin kinase.

Publication ,  Journal Article
Inglese, J; Glickman, JF; Lorenz, W; Caron, MG; Lefkowitz, RJ
Published in: J Biol Chem
January 25, 1992

The primary structure of bovine rhodopsin kinase (RK), which phosphorylates light-activated rhodopsin (Rho*), terminates with the amino acid sequence Cys558-Val-Leu-Ser561, a motif that has been shown to direct the isoprenylation and alpha-carboxyl methylation of many proteins (e.g. p21Ha-ras). Transient expression of RK in COS-7 cells revealed the presence of two immunoreactive protein species. Consistent with RK being modified by isoprenylation, interconversion of these two species was dependent upon isoprenoid biosynthesis in the cells. Moreover, a serine substitution for Cys558 resulted in a single RK species whose migration on sodium dodecyl sulfate-polyacrylamide gels was identical to that of RK from cells treated with mevinolin, an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase and, thus, of isoprenoid biosynthesis. This finding indicates that isoprenylation of RK requires Cys558. The electrophoretic mobility of isoprenylated RK synthesized in COS-7 cells was identical to that of RK from bovine rod outer segments, suggesting that RK is isoprenylated in vivo. RK was determined to be modified by a farnesyl moiety and alpha-carboxyl-methylated. A time course of Rho* phosphorylation revealed that non-processed RK is approximately 4-fold less active than wild-type RK. This is the first demonstration of isoprenylation/alpha-carboxyl methylation of a protein kinase, and suggests that these modifications markedly influence enzymatic activity in vivo.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

January 25, 1992

Volume

267

Issue

3

Start / End Page

1422 / 1425

Location

United States

Related Subject Headings

  • Transfection
  • Rod Cell Outer Segment
  • Recombinant Fusion Proteins
  • Protein Processing, Post-Translational
  • Protein Kinases
  • Mutagenesis, Site-Directed
  • Molecular Weight
  • Molecular Sequence Data
  • Mevalonic Acid
  • Methylation
 

Citation

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Inglese, J., Glickman, J. F., Lorenz, W., Caron, M. G., & Lefkowitz, R. J. (1992). Isoprenylation of a protein kinase. Requirement of farnesylation/alpha-carboxyl methylation for full enzymatic activity of rhodopsin kinase. J Biol Chem, 267(3), 1422–1425.
Inglese, J., J. F. Glickman, W. Lorenz, M. G. Caron, and R. J. Lefkowitz. “Isoprenylation of a protein kinase. Requirement of farnesylation/alpha-carboxyl methylation for full enzymatic activity of rhodopsin kinase.J Biol Chem 267, no. 3 (January 25, 1992): 1422–25.
Inglese J, Glickman JF, Lorenz W, Caron MG, Lefkowitz RJ. Isoprenylation of a protein kinase. Requirement of farnesylation/alpha-carboxyl methylation for full enzymatic activity of rhodopsin kinase. J Biol Chem. 1992 Jan 25;267(3):1422–5.
Inglese J, Glickman JF, Lorenz W, Caron MG, Lefkowitz RJ. Isoprenylation of a protein kinase. Requirement of farnesylation/alpha-carboxyl methylation for full enzymatic activity of rhodopsin kinase. J Biol Chem. 1992 Jan 25;267(3):1422–1425.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

January 25, 1992

Volume

267

Issue

3

Start / End Page

1422 / 1425

Location

United States

Related Subject Headings

  • Transfection
  • Rod Cell Outer Segment
  • Recombinant Fusion Proteins
  • Protein Processing, Post-Translational
  • Protein Kinases
  • Mutagenesis, Site-Directed
  • Molecular Weight
  • Molecular Sequence Data
  • Mevalonic Acid
  • Methylation