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Phosphorylation of the beta-adrenergic receptor in intact cells: relationship to heterologous and homologous mechanisms of adenylate cyclase desensitization.

Publication ,  Journal Article
Sibley, DR; Daniel, K; Strader, CD; Lefkowitz, RJ
Published in: Arch Biochem Biophys
October 1987

We have recently shown that both heterologous and homologous forms of adenylate cyclase desensitization involve phosphorylation of beta-adrenergic receptors. In order to compare these two reactions, we wished to identify a single cell system in which both processes could be studied. Using the frog erythrocyte, which has been previously shown to exhibit cAMP-independent homologous desensitization, we have found that under appropriate conditions cAMP-dependent heterologous desensitization can be elicited. Incubation of intact cells with the membrane-permeable cAMP analogs dibutyryl cAMP or 8-bromo cAMP promotes about a 50% desensitization of isoproterenol- and prostaglandin E1-stimulated adenylate cyclase activity in a time-, temperature-, and dose-dependent fashion. There is also a 20% desensitization in the abilities of guanine nucleotides (GTP and guanyl-5'-yl-imidodiphosphate) and NaF to stimulate adenylate cyclase maximally. In contrast, there is no effect on forskolin- or MnCl2-stimulated enzyme activities. The desensitization response is specific for cAMP as dibutyryl cGMP, 8-bromo cGMP, or 8-bromo AMP produce little or no desensitization. Incubation of the cells with dibutyryl cAMP does not affect the number of cell surface beta-adrenergic receptors. In contrast, incubation with isoproterenol promotes homologous desensitization and sequestration of the receptors. Incubation of 32P-labeled erythrocytes with either dibutyryl cAMP or isoproterenol promotes a stoichiometric threefold increase in the phosphorylation state of the beta-adrenergic receptor which occurs predominantly on serine residues. However, if the cells are coincubated with both dibutyryl cAMP and isoproterenol then the desensitization of isoproterenol-stimulated enzyme activity and phosphorylation of the beta-adrenergic receptor are greater than those observed with either agent alone. These results indicate that heterologous and homologous desensitization of adenylate cyclase-coupled beta-adrenergic receptors are mediated by different biochemical pathways involving phosphorylation of the receptor protein on distinct sites.

Duke Scholars

Published In

Arch Biochem Biophys

DOI

ISSN

0003-9861

Publication Date

October 1987

Volume

258

Issue

1

Start / End Page

24 / 32

Location

United States

Related Subject Headings

  • Sodium Fluoride
  • Receptors, Adrenergic, beta
  • Rana pipiens
  • Phosphorylation
  • Isoproterenol
  • Guanylyl Imidodiphosphate
  • Guanosine Triphosphate
  • Erythrocytes
  • Electrophoresis, Polyacrylamide Gel
  • Cyclic AMP
 

Citation

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Sibley, D. R., Daniel, K., Strader, C. D., & Lefkowitz, R. J. (1987). Phosphorylation of the beta-adrenergic receptor in intact cells: relationship to heterologous and homologous mechanisms of adenylate cyclase desensitization. Arch Biochem Biophys, 258(1), 24–32. https://doi.org/10.1016/0003-9861(87)90318-3
Sibley, D. R., K. Daniel, C. D. Strader, and R. J. Lefkowitz. “Phosphorylation of the beta-adrenergic receptor in intact cells: relationship to heterologous and homologous mechanisms of adenylate cyclase desensitization.Arch Biochem Biophys 258, no. 1 (October 1987): 24–32. https://doi.org/10.1016/0003-9861(87)90318-3.
Sibley, D. R., et al. “Phosphorylation of the beta-adrenergic receptor in intact cells: relationship to heterologous and homologous mechanisms of adenylate cyclase desensitization.Arch Biochem Biophys, vol. 258, no. 1, Oct. 1987, pp. 24–32. Pubmed, doi:10.1016/0003-9861(87)90318-3.
Journal cover image

Published In

Arch Biochem Biophys

DOI

ISSN

0003-9861

Publication Date

October 1987

Volume

258

Issue

1

Start / End Page

24 / 32

Location

United States

Related Subject Headings

  • Sodium Fluoride
  • Receptors, Adrenergic, beta
  • Rana pipiens
  • Phosphorylation
  • Isoproterenol
  • Guanylyl Imidodiphosphate
  • Guanosine Triphosphate
  • Erythrocytes
  • Electrophoresis, Polyacrylamide Gel
  • Cyclic AMP