Serotonin-induced protein kinase C activation in cultured rat heart endothelial cells.
This study examined whether serotonin can activate protein kinase C in rat heart endothelial cells. Protein kinase C isozyme translocation was examined by Western blot analysis with isozyme-specific anti-protein kinase C antibody. In this study, only alpha protein kinase C isozyme was found to be translocated from the cytosolic to the particulate fractions after serotonin stimulation. The effect of serotonin on the incorporation of 32P from [gamma-32P]ATP into peptide substrate was studied as another indicator of protein kinase C activation. The experiments in this study demonstrated that the Ca(2+)-phospholipid-dependent protein kinase, protein kinase C, was activated by serotonin. By investigating [3H]phorbol 12,13-dibutyrate binding to protein kinase C and trypsin-treated protein kinase C activity, we demonstrated that the site of action of serotonin is probably the regulatory domain of protein kinase C. Finally, we also demonstrated that serotonin had no effect on the intracellular concentration of cyclic nucleotides (cAMP, cGMP). These findings support the hypothesis that protein kinase C may be an important participant in serotonin-induced endothelial cell contraction and barrier dysfunction.
Lee
H Z
HZ
School of Pharmacy, China Medical College, 91, Hsueh-Shih Road, 404, Taichung, Taiwan. hong@mail.cmc.edu.tw
Wu
C
C
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009206:Myocardium / Q000166:cytology / Q000201:enzymology*; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D012701:Serotonin / Q000494:pharmacology*; D014178:Translocation, Genetic / Q000187:drug effects; D014357:Trypsin / Q000494:pharmacology
D012701:Serotonin; D015240:Phorbol 12,13-Dibutyrate; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D007770:L-Lactate Dehydrogenase; D011493:Protein Kinase C; D014357:Trypsin; D006152:Cyclic GMP