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 phospholipase D and whether its enzymatic activity is stimulated by phorbol-12-myristate-13-acetate, Sar super(1)-angiotensin II, or bradykinin. Phospholipase D immunoreactivity was detected in the cultured cells by Western blot, and it was mainly found in the membrane protein extract. In cells labelled with [ super(3)H]oleic acid and exposed to butanol and the stimulants, phorbol-12-myristate-13-acetate (1 mu M) markedly increased the accumulation of [ super(3)H]phosphatidylbutanol in five minutes, and the stimulatory effect was sustained for one hour. Angiotensin (200 nM) and bradykinin (100 nM) also increased the accumulation of phosphatidylbutanol but their effect was not sustained. We conclude that bovine adrenal chromaffin cells have phospholipase D activity that is stimulated by angiotensin II, bradykinin, and activation of protein kinase C.