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Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism.

Publication ,  Journal Article
Leeb-Lundberg, LM; Cotecchia, S; Lomasney, JW; DeBernardis, JF; Lefkowitz, RJ; Caron, MG
Published in: Proc Natl Acad Sci U S A
September 1985

DDT1 MF-2 cells, which are derived from hamster vas deferens smooth muscle, contain alpha 1-adrenergic receptors (54,800 +/- 2700 sites per cell) that are coupled to stimulation of inositol phospholipid metabolism. Incubation of these cells with tumor-promoting phorbol esters, which stimulate calcium- and phospholipid-dependent protein kinase, leads to a marked attenuation of the ability of alpha 1-receptor agonists such as norepinephrine to stimulate the turnover of inositol phospholipids. This turnover was measured by determining the 32P content of phosphatidylinositol and phosphatidic acid after prelabeling of the cellular ATP pool with 32Pi. These phorbol ester-treated cells also displayed a decrease in binding affinity of cellular alpha 1 receptors for agonists with no change in antagonist affinity. By using affinity chromatography on the affinity resin Affi-Gel-A55414, the alpha 1 receptors were purified approximately equal to 300-fold from control and phorbol ester-treated 32Pi-prelabeled cells. As assessed by NaDodSO4/polyacrylamide gel electrophoresis, the Mr 80,000 alpha 1-receptor ligand-binding subunit is a phosphopeptide containing 1.2 mol of phosphate per mol of alpha 1 receptor. After phorbol ester treatment this increased to 3.6 mol of phosphate per mol of alpha 1 receptor. The effect of phorbol esters on norepinephrine-stimulated inositol phospholipid turnover and alpha 1-receptor phosphorylation showed the same rapid time course with a t1/2 less than 2 min. These results indicate that calcium- and phospholipid-dependent protein kinase may play an important role in regulating the function of receptors that are coupled to the inositol phospholipid cycle by phosphorylating and deactivating them.

Duke Scholars

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

September 1985

Volume

82

Issue

17

Start / End Page

5651 / 5655

Location

United States

Related Subject Headings

  • Vas Deferens
  • Tetradecanoylphorbol Acetate
  • Receptors, Adrenergic, alpha
  • Protein Kinases
  • Protein Kinase C
  • Phosphorylation
  • Phosphoproteins
  • Phosphatidylinositols
  • Phorbols
  • Norepinephrine
 

Citation

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MLA
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Leeb-Lundberg, L. M., Cotecchia, S., Lomasney, J. W., DeBernardis, J. F., Lefkowitz, R. J., & Caron, M. G. (1985). Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism. Proc Natl Acad Sci U S A, 82(17), 5651–5655. https://doi.org/10.1073/pnas.82.17.5651
Leeb-Lundberg, L. M., S. Cotecchia, J. W. Lomasney, J. F. DeBernardis, R. J. Lefkowitz, and M. G. Caron. “Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism.Proc Natl Acad Sci U S A 82, no. 17 (September 1985): 5651–55. https://doi.org/10.1073/pnas.82.17.5651.
Leeb-Lundberg LM, Cotecchia S, Lomasney JW, DeBernardis JF, Lefkowitz RJ, Caron MG. Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5651–5.
Leeb-Lundberg, L. M., et al. “Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism.Proc Natl Acad Sci U S A, vol. 82, no. 17, Sept. 1985, pp. 5651–55. Pubmed, doi:10.1073/pnas.82.17.5651.
Leeb-Lundberg LM, Cotecchia S, Lomasney JW, DeBernardis JF, Lefkowitz RJ, Caron MG. Phorbol esters promote alpha 1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5651–5655.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

September 1985

Volume

82

Issue

17

Start / End Page

5651 / 5655

Location

United States

Related Subject Headings

  • Vas Deferens
  • Tetradecanoylphorbol Acetate
  • Receptors, Adrenergic, alpha
  • Protein Kinases
  • Protein Kinase C
  • Phosphorylation
  • Phosphoproteins
  • Phosphatidylinositols
  • Phorbols
  • Norepinephrine