Enhancement of bradykinin-induced prostacyclin synthesis in porcine aortic endothelial cells by pertussis toxin. Possible implication of lipocortin I.
Bradykinin-stimulated prostacyclin synthesis in porcine aortic endothelial cells was enhanced by pretreatment of the cells with pertussis toxin or islet-activating protein (IAP) for 5 hr or longer. Although ADP-ribosylation of a protein with a molecular weight of 41-42 kD in the cell membranes was completed by 3 hr after the addition of IAP into the incubation medium, there was good correlation between enhancement of bradykinin-induced prostacyclin synthesis and ADP-ribosylation of the IAP substrate over a wide range of IAP concentrations. Furthermore, even if IAP was removed from the incubation medium at 3 hr, bradykinin-induced prostaglandin synthesis at 24 hr was still potentiated. Cycloheximide and actinomycin D enhanced bradykinin-induced prostacyclin synthesis and apparently blocked the effect of IAP. Since this result suggested the involvement of an inhibitor protein(s) of prostacyclin synthesis in the IAP effect, we studied the effect of IAP on the level of lipocortin I which is known to inhibit phospholipase A2. Western and Northern blot analyses revealed that IAP decreased the amounts of protein and mRNA of lipocortin I. These results suggest that the enhancement of bradykinin-induced prostacyclin synthesis by IAP is associated with a decrease in the level of lipocortin I.
Fujimoto
M
M
Shionogi Research Laboratories, Shionogi & Co., Ltd., Osaka, Japan.
Sakata
T
T
Tsuruta
Y
Y
Iwagami
S
S
Teraoka
H
H
Mihara
S
S
Fukiishi
Y
Y
Ide
M
M
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D017302:Annexins; D001011:Aorta; D015152:Blotting, Northern; D001920:Bradykinin / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000502:physiology*; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000378:metabolism*; D011464:Epoprostenol / Q000096:biosynthesis*; D015870:Gene Expression / Q000187:drug effects; D007295:Inositol Phosphates / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight; D009355:Neomycin / Q000494:pharmacology; D037342:Pertussis Toxin; D012333:RNA, Messenger / Q000235:genetics; D013552:Swine; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D017302:Annexins; D002135:Calcium-Binding Proteins; D007295:Inositol Phosphates; D008565:Membrane Proteins; D012333:RNA, Messenger; D010566:Virulence Factors, Bordetella; D003609:Dactinomycin; D000246:Adenosine Diphosphate Ribose; D003513:Cycloheximide; D011464:Epoprostenol; D037342:Pertussis Toxin; D009355:Neomycin; D001920:Bradykinin