title: Interactions of bradykinin, calcium, G-protein and protein kinase in the activation of phospholipase A2 in bovine pulmonary artery endothelial cells.
abstract: Rise in free cytosolic calcium concentrations [Ca2+]i in response to bradykinin and guanosine 5'-O-thiotriphosphate (GTP tau S) was related to the action of phospholipase A/ (arachidonic acid release). At 900 microM extracellular CaCl2, bradykinin induced a typical Ca2+ movement consisting of an initial [Ca2+]i peak at approximately 400 nM followed by a sustained increase in the steady-state cytosolic Ca2+ level at approximately 290 nM. As the extracellular CaCl2 concentration was reduced to 100 microM, the bradykinin induced initial spike was reduced followed by only a marginal increase in steady-state cytosolic Ca2+ levels. Treatment of endothelial cells with saponin (0.002% w/w) did not increase [Ca2+]i and saponin treated cells exhibited a very similar pattern of Ca2+ mobilization in response to bradykinin. However, with saponin treatment, GTP tau S (100 microM) increased [Ca2+]i at an almost identical tracing exhibited with 50 nM bradykinin stimulation (in either the presence or absence of 0.002% saponin). No additive increase in [Ca2+]i was observed in cells stimulated with both 100 microM GTP tau S and 50 nM bradykinin or in bradykinin stimulated cells subsequently exposed to GTP tau S. Pertussis toxin (PTX) did not affect the bradykinin induced Ca2+ mobilization. However, as we showed previously, PTX inhibited bradykinin stimulated arachidonic acid release. These results indicate transduction of the bradykinin signal by G-protein for both phospholipase A2 (PLA2) activation and Ca2+ mobilization but likely by different G alpha subunits, a PTX sensitive and an insensitive subunit. Furthermore, the bradykinin and GTP tau S stimulated release of arachidonic acid appears to be only partially dependent on [Ca2+]i.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Boston University School of Medicine, MA 02118.
authors.forename: D
authors.initials: D
authors.lastname: Ricupero
authors.forename: L
authors.initials: L
authors.lastname: Taylor
authors.forename: P
authors.initials: P
authors.lastname: Polgar
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001920:Bradykinin / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism* / Q000502:physiology; D002417:Cattle; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000031:analogs & derivatives / Q000494:pharmacology; D017736:Ion-Selective Electrodes; D037342:Pertussis Toxin; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011494:Protein Kinases / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000187:drug effects / Q000201:enzymology; D012503:Saponins / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
chemical_list: D012503:Saponins; D010566:Virulence Factors, Bordetella; C034882:guanosine 5'-O-(1-thiotriphosphate); D016718:Arachidonic Acid; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D037342:Pertussis Toxin; D011494:Protein Kinases; D011493:Protein Kinase C; D010741:Phospholipases A; D054467:Phospholipases A2; D019204:GTP-Binding Proteins; D013755:Tetradecanoylphorbol Acetate; D001920:Bradykinin; D002118:Calcium