Phorbol esters promote α1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism

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Phorbol esters promote α1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism. / Leeb-Lundberg, L. M.; Cotecchia, S.; Lomasney, J. W.; DeBernardis, J. F.; Lefkowitz, R. J.; Caron, M. G.

I: Proceedings of the National Academy of Sciences of the United States of America, Vol. 82, Nr. 17, 01.12.1985, s. 5651-5655.

Forskningsoutput: TidskriftsbidragArtikel i vetenskaplig tidskrift

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T1 - Phorbol esters promote α1-adrenergic receptor phosphorylation and receptor uncoupling from inositol phospholipid metabolism

AU - Leeb-Lundberg, L. M.

AU - Cotecchia, S.

AU - Lomasney, J. W.

AU - DeBernardis, J. F.

AU - Lefkowitz, R. J.

AU - Caron, M. G.

PY - 1985/12/1

Y1 - 1985/12/1

N2 - DDT1 MF-2 cells, which are derived from hamster vas deferens smooth muscle, contain α1-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 α1-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 32P(i). These phorbol ester-treated cells also displayed a decrease in binding affinity of cellular α1 receptors for agonists with no change in antagonist affinity. By using affinity chromatography on the affinity resin Affi-Gel-A55414, the α1 receptors were purified ~300-fold from control and phorbol ester-treated 32P(i)-prelabeled cells. As assessed by NaDodSO4/polyacrylamide gel electrophoresis, the M(r) 80,000 α1-receptor ligand-binding subunit is a phosphopeptide containing 1.2 mol of phosphate per mol of α1 receptor. After phorbol ester treatment this increased to 3.6 mol of phosphate per mol of α1 receptor. The effect of phorbol esters on norepinephrine-stimulated inositol phospholipid turnover and α1-receptor phosphorylation showed the same rapid time course with a t( 1/2 )

AB - DDT1 MF-2 cells, which are derived from hamster vas deferens smooth muscle, contain α1-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 α1-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 32P(i). These phorbol ester-treated cells also displayed a decrease in binding affinity of cellular α1 receptors for agonists with no change in antagonist affinity. By using affinity chromatography on the affinity resin Affi-Gel-A55414, the α1 receptors were purified ~300-fold from control and phorbol ester-treated 32P(i)-prelabeled cells. As assessed by NaDodSO4/polyacrylamide gel electrophoresis, the M(r) 80,000 α1-receptor ligand-binding subunit is a phosphopeptide containing 1.2 mol of phosphate per mol of α1 receptor. After phorbol ester treatment this increased to 3.6 mol of phosphate per mol of α1 receptor. The effect of phorbol esters on norepinephrine-stimulated inositol phospholipid turnover and α1-receptor phosphorylation showed the same rapid time course with a t( 1/2 )

U2 - 10.1073/pnas.82.17.5651

DO - 10.1073/pnas.82.17.5651

M3 - Article

VL - 82

SP - 5651

EP - 5655

JO - Proceedings of the National Academy of Sciences

T2 - Proceedings of the National Academy of Sciences

JF - Proceedings of the National Academy of Sciences

SN - 1091-6490

IS - 17

ER -