Phorbol diesters promote β-adrenergic receptor phosphorylation and adenylate cyclase desensitization in duck erythrocytes

Sibley, David R.; Nambi, Ponnal; Peters, J R; Lefkowiz, Robert J.

doi:10.1016/0006-291x(84)90772-1

Cited by 229 publications

(32 citation statements)

References 14 publications

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“…insulin (Jacobs et al, 1983;Takayama et al, 1984), a1-adrenoceptor (Leeb-Lundberg et al, 1985) and #-adrenoceptor (Sibley et al, 1984;Kelleher et al, 1984), and that in some of these systems this is associated with a reduction in agonist binding affinity and in response (see for example Leeb-Lundberg et al, 1985).…”

Section: Discussionmentioning

confidence: 99%

Release of endothelium‐derived relaxing factor from pig cultured aortic endothelial cells, as assessed by changes in endothelial cell cyclic GMP content, is inhibited by a phorbol ester

Smith

Lang

1990

British J Pharmacology

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Cultured aortic endothelial cells of the pig respond to the endothelium‐derived relaxing factor (EDRF) they release with an increase in cyclic GMP content. This response is inhibited by haemoglobin or by l‐NG‐monomethyl‐arginine (l‐NMMA), and has been used to investigate the effects of phorbol esters on EDRF release. Pretreatment with phorbol‐12,13‐dibutyrate (PDB) but not the inactive 4α‐phorbol‐12,13,‐didecanoate (PDD), inhibited increases in cyclic GMP induced by substance P (10−8 m) in a time and concentration‐dependent manner. PDB did not affect basal cyclic GMP levels. PDB (3 × 10−7 m), but not PDD (3 × 10−7 m), also inhibited ATP (10−5 m)‐induced increases in cyclic GMP, but did not affect those induced by bradykinin (10−7 m). Increases in cyclic GMP induced by low (10−7 m) but not high (10−6 m) concentrations of the calcium ionophore A23187 were inhibited by PDB (3 × 10−7 m). This inhibitory effect was due to enhanced destruction of EDRF by superoxide anions rather than inhibition of EDRF release, as the inhibition was abolished in the presence of superoxide dismutase (SOD, 30 u ml−1) and catalase (CAT, 100 u ml−1). SOD and CAT did not affect the inhibitory action of PDB on substance P or ATP‐induced increases in cyclic GMP. Increases in endothelial cell cyclic GMP content induced by sodium nitroprusside (10−5 m) were unaffected by PDB pretreatment. The inhibitory effects of PDB are probably a result of an action of protein kinase C on the steps between receptor occupation and phospholipase C activation.

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Section: Discussionmentioning

confidence: 99%

Release of endothelium‐derived relaxing factor from pig cultured aortic endothelial cells, as assessed by changes in endothelial cell cyclic GMP content, is inhibited by a phorbol ester

Smith

Lang

1990

British J Pharmacology

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“…Moreover, the biochemical signal transfer pathways for these receptors have not yet been clearly elucidated. The adenylate cyclasecoupled ,3-adrenergic receptors have also been shown to be a target of protein kinase C-mediated phosphorylation (27,28). This covalent modification of the f3 receptor appears to uncouple it from adenylate cyclase stimulation, thus leading to "desensitization" of the receptor response.…”

mentioning

confidence: 99%

“…In each case the phosphorylated receptor appears to be functionally uncoupled from its biological effector system (27,28 …”

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confidence: 99%

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

Leeb-Lundberg¹,

Cotecchia²,

Lomasney³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

284

118

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DDT, MF-2 cells, which are derived from hamster vas deferens smooth muscle, contain a1-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 a,-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;. These phorbol ester-treated cells also displayed a decrease in binding affinity of cellular al receptors for agonists with no change in antagonist affinity. By using affinity chromatography on the affinity resin Affi-Gel-A55414, the a, receptors were purified =300-fold from control and phorbol ester-treated 32P1-prelabeled cells. As assessed by NaDodSO4/polyacrylamide gel electrophoresis, the Mr 80,000 a,-receptor ligandbinding subunit is a phosphopeptide containing 1.2 mol of phosphate per mol of a, receptor. After phorbol ester treatment this increased to 3.6 mol of phosphate per mol of a, receptor. The effect of phorbol esters on norepinephrinestimulated inositol phospholipid turnover and a,-receptor phosphorylation showed the same rapid time course with a til2 < 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.The mechanisms whereby hormones regulate cellular metabolism are of great current biochemical interest. In addition to the well-delineated, second messenger-generating adenylate cyclase system, another potentially quite general pathway has been demonstrated recently. This pathway involves receptor-mediated stimulation of the breakdown of membrane polyphosphoinositides (phosphatidylinositol phosphates) to diacylglycerol and inositol trisphosphate (1-5).Both products are potential second messengers serving to stimulate calcium-and phospholipid-dependent protein kinase (protein kinase C) (6, 7) and to mobilize intracellular calcium (8, 9), respectively. Factors known to stimulate this pathway include agonists such as acetylcholine (10), angiotensin (11), and catecholamines acting via a1-adrenergic receptors (1,(12)(13)(14)(15)(16).A powerful approach to the study of this signal transfer pathway has been the use of the tumor-promoting phorbol esters, which appear to mimic endogenously produced diacylglycerol in activating protein kinase C (17, 18), thus bypassing the receptor signal. By using this approach it has been demonstrated recently that receptors for epidermal growth factor (19, 20), insulin (21, 22), and somatomedin C (21) are either directly or indirectly phosphorylated by protein kinase C. Although in a few of these systems receptor phosphorylation has been shown to...

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“…Then, all the inhibitory input activities which would stimulate Gk to produce hyperpolarization in the neuron could be suppressed because protein kinase C would deactivate Gk and K+-channel activities. In contrast, the excitatory input activities which would stimulate GS to produce depolarization in the same neuron could be augmented if protein kinase C is mildly activated just to phosphorylate and deactivate G; [57]. However, they could be rather depressed if protein kinase C is activated intensely to phosphorylate the receptor protein itself [58].…”

Section: Neurophysiological Relevancementioning

confidence: 98%

GTP-binding proteins and their regulatory actions on ion channels.

Sato

1989

Jpn.J.Physiol

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Phorbol diesters promote β-adrenergic receptor phosphorylation and adenylate cyclase desensitization in duck erythrocytes

Cited by 229 publications

References 14 publications

Release of endothelium‐derived relaxing factor from pig cultured aortic endothelial cells, as assessed by changes in endothelial cell cyclic GMP content, is inhibited by a phorbol ester

Release of endothelium‐derived relaxing factor from pig cultured aortic endothelial cells, as assessed by changes in endothelial cell cyclic GMP content, is inhibited by a phorbol ester

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

GTP-binding proteins and their regulatory actions on ion channels.

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