Alpha 1-adrenergic receptors promote phosphatidylcholine hydrolysis in MDCK-D1 cells. A mechanism for rapid activation of protein kinase C.
Hydrolysis of phosphoinositides can lead to mobilization of calcium and production of diacylglycerol, which together are proposed to activate protein kinase C. We have shown that phosphoinositide hydrolysis mediated by alpha 1-adrenergic receptors on Madin-Darby canine kidney cells (MDCK-D1) occurred with an early lag and increased over a prolonged time course (Slivka, S.R., and Insel, P.A. (1987) J. Biol. Chem. 262, 4200-4207). In this study we characterize another type of receptor-mediated phospholipid hydrolysis in MDCK-D1 cells, alpha 1-adrenergic receptor-mediated hydrolysis of phosphatidylcholine. The predicted products of this hydrolysis, phosphorylcholine and diacylglycerol, were detectable as early as 0.5 min after alpha 1-adrenergic receptor stimulation by epinephrine. This hydrolysis appears to be a primary event after receptor occupancy because it occurred in the presence of neomycin, an inhibitor of polyphosphoinositide hydrolysis, and the protein kinase C inhibitors, sphingosine and 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7). In addition, we demonstrate alpha 1-adrenergic receptor-mediated activation of protein kinase C in MDCK-D1 cells. This activation was measured as a rapid translocation (0.5 min) of protein kinase C activity from the cytosolic fraction to the membrane fraction. This translocation also was not inhibited by neomycin. The time course and agonist concentration dependence of both phosphatidylcholine hydrolysis and protein kinase C activation by alpha 1-adrenergic receptors were similar. Thus, we propose that agonists acting at alpha 1-adrenergic receptors promote hydrolysis of phosphatidylcholine which results in rapid generation of diacylglycerol for the activation of protein kinase C.
Slivka
S R
SR
Department of Pharmacology, University of California, San Diego, La Jolla 92093.
Meier
K E
KE
Insel
P A
PA
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D003600:Cytosol / Q000201:enzymology; D004075:Diglycerides / Q000378:metabolism; D004285:Dogs; D004789:Enzyme Activation; D004837:Epinephrine / Q000494:pharmacology; D006868:Hydrolysis; D007546:Isoquinolines / Q000494:pharmacology; D007668:Kidney; D007700:Kinetics; D009355:Neomycin / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000378:metabolism*; D010767:Phosphorylcholine / Q000378:metabolism; D010879:Piperazines / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011942:Receptors, Adrenergic, alpha / Q000187:drug effects / Q000502:physiology*; D013110:Sphingosine / Q000494:pharmacology
D004075:Diglycerides; D007546:Isoquinolines; D010713:Phosphatidylcholines; D010879:Piperazines; D011942:Receptors, Adrenergic, alpha; D010767:Phosphorylcholine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; D009355:Neomycin; D013110:Sphingosine; D004837:Epinephrine