Effect of inhibitors of mitogen-activated protein kinase kinase on alpha(1B)-adrenoceptor phosphorylation.
1 Mitogen-activated protein kinases mediate hormone/neurotransmitter action on proliferation and differentiation and participate in receptor regulation. The effect of inhibitors of mitogen-activated kinase kinase (MEK) on alpha(1B)-adrenoceptor phosphorylation state and function was studied using different cell lines. It was observed that at nanomolar concentrations the MEK inhibitors, PD98059 (2'-amino-3'-methoxyflavone) and UO126 [1,4-(diamino-2,3-dicyano/1,4-bis-(2-aminophenylthio)-butadiene], increased alpha(1B)-adrenoceptor phosphorylation and diminished the functional response of this receptor to noradrenaline. These agents did not alter the action of lysophosphatidic acid. 2 Staurosporine (IC(50) approximately 0.8 nm) (a general protein kinase inhibitor) and bis-indolyl-maleimide I (IC(50) approximately 200 nm) (a selective protein kinase C inhibitor) inhibited PD98059-induced alpha(1B)-adrenoceptor phosphorylation. In contrast, neither wortmannin (phosphoinositide 3-kinase inhibitor) nor genistein (protein tyrosine kinase inhibitor) had any effect. The data suggest the possibility that MEK might exert control on the activity of the enzymes that regulate receptor phosphorylation, such as G-protein-coupled receptor kinases, protein kinase C or serine/threonine protein phosphatases. 3 Coimmunoprecipitation studies showed a constant association of total extracellular signal-regulated kinase 2 (ERK2) with alpha(1B)-adrenoceptors. Association of phospho-ERK 1/2 to alpha(1B)-adrenoceptors increased not only in response to agonist but also in response to agents that increase alpha(1B)-adrenoceptor and ERK1/2 phosphorylation [such as endothelin-1, phorbol 12-myristate-13-acetate (PMA) and epidermal growth factor (EGF)]; not surprisingly, PD98059 decreased this effect. 4 Our data show that blockade of MEK activity results in increased alpha(1B)-adrenoceptor phosphorylation, diminished adrenoceptor function and perturbation of receptor-ERK1/2 interaction.
Alcántara-Hernández
R
R
Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, D.F., Mexico.
Adolfo García-Sáinz
J
J
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D002070:Butadienes / Q000494:pharmacology; D020013:Calcium Signaling / Q000187:drug effects; D002460:Cell Line; D006224:Cricetinae; D019332:Endothelin-1 / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008246:Lysophospholipids / Q000494:pharmacology; D048370:MAP Kinase Kinase 2 / Q000235:genetics / Q000378:metabolism; D008301:Maleimides / Q000494:pharmacology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D009638:Norepinephrine / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D000077191:Wortmannin
C547716:ADRA1B protein, human; D000730:Androstadienes; D002070:Butadienes; D019332:Endothelin-1; D005419:Flavonoids; D007211:Indoles; D008246:Lysophospholipids; D008301:Maleimides; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D034741:RNA, Small Interfering; D018340:Receptors, Adrenergic, alpha-1; C113580:U 0126; D004815:Epidermal Growth Factor; D019833:Genistein; C482128:MAP2K2 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048370:MAP Kinase Kinase 2; D020929:Mitogen-Activated Protein Kinase Kinases; D019311:Staurosporine; C088060:bisindolylmaleimide; D013755:Tetradecanoylphorbol Acetate; C032881:lysophosphatidic acid; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D009638:Norepinephrine; D000077191:Wortmannin