Lysophosphatidic acid modulates alpha(1b)-adrenoceptor phosphorylation and function: roles of Gi and phosphoinositide 3-kinase.
The effect of lysophosphatidic acid on the phosphorylation and function of alpha(1b)-adrenoceptors transfected into rat-1 fibroblasts was studied. This phospholipid mitogen increased in a concentration-dependent fashion (EC(50) approximately 50 nM) the phosphorylation of these adrenoceptors. Lysophosphatidic acid-induced alpha(1b)-adrenoceptor phosphorylation was relatively rapid (t(1/2) approximately 1 min), intense (2.5-fold), and sustained for at least 60 min. The effect of lysophosphatidic acid was blocked by pretreatment with pertussis toxin. The alpha(1b)-adrenoceptor phosphorylation induced by lysophosphatidic acid was not blocked by genistein, a tyrosine kinase inhibitor, but it was inhibited by inhibitors of protein kinase C (bisindolylmaleimide I, staurosporine, and Ro 31-8220) and phosphoinositide 3-kinase (wortmannin and LY 294002). The ability of norepinephrine to increase cytosol calcium concentration was markedly decreased in cells previously challenged with lysophosphatidic acid. Norepinephrine-induced [(35)S]GTPgammaS binding in membrane preparations was used as an index of the functional coupling of the alpha(1b)-adrenoceptors and G proteins. Norepinephrine-stimulated [(35)S]GTPgammaS binding was markedly decreased in membranes from cells pretreated with lysophosphatidic acid. This effect of lysophosphatidic acid was blocked by pretreatment with wortmannin or staurosporine. Our data indicate that: 1) activation of lysophosphatidic acid receptors induce phosphorylation of alpha(1b)-adrenoceptors; 2) this effect is mediated through pertussis toxin-sensitive G proteins, phosphatidylinositol 3-kinase, and protein kinase C; and 3) the phosphorylation of alpha(1b)-adrenoceptors induced by the lipid mitogen is associated to adrenoceptor desensitization.
Casas-González
P
P
Departamento de Biología Celular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México City, México.
Vázquez-Prado
J
J
García-Sáinz
J A
JA
D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000502:physiology*; D008246:Lysophospholipids / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000502:physiology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000502:physiology; D051381:Rats; D018340:Receptors, Adrenergic, alpha-1 / Q000378:metabolism* / Q000502:physiology
C547718:Adra1b protein, rat; D008246:Lysophospholipids; D018340:Receptors, Adrenergic, alpha-1; D019869:Phosphatidylinositol 3-Kinases; D011493:Protein Kinase C; D019206:GTP-Binding Protein alpha Subunits, Gi-Go