Receptor-stimulated phospholipase D activity in bovine adrenal chromaffin cells

Bovine adrenal chromaffin cells have been thought to lack phospholipase D. We reinvestigated whether primary cultured bovine adrenal chromaffin cells possess phospholipass D and whether its enzymatic activity is stimulated by phorbol-12-myristate-13-acetate, Sap(1)-angiotensin II, or bradykinin. Pho spholipase D immunoreactivity was detected in the cultured cells by Western blot, and it was mainly found in the membrane protein extract. In cells la belled with [H-3]oleic acid and exposed to butanol and the stimulants, phor bol-12-myristate-13-acetate (1 mu M) markedly increased the accumulation of [H-3]phosphatidylbutanol in five minutes, and the stimulatory effect was s ustained for one hour. Angiotensin (200 nM) and bradykinin (100 nM) also in creased the accumulation of phosphatidylbutanol but their effect was not su stained. We conclude that bovine adrenal chromaffin cells have phospholipas e D activity that is stimulated by angiotensin II, bradykinin, and activati on of protein kinase C.