Protein kinase C-mediated inhibition of renal Ca2+ ATPase by physiological concentrations of angiotensin II is reversed by AT1- and AT2-receptor antagonists

Angiotensin II (Ang II) increases the cytosolic Ca2+ concentration in different cell types. In this study, we investigate the effect of Ang II on the Ca2+ ATPase of purified basolateral membranes of kidney proximal tubules. This enzyme pumps Ca2+ out of the cytosol in a reaction coupled to ATP hydrolysis, and it is responsible for the fine-timed regulation of cytosolic Ca2+ activity. Ca2+-ATPase activity is inhibited by picomolar concentrations of Ang II, with maximal inhibition being attained at approximate to 50% of the control values. The presence of raising concentrations (10(-11) to 10(-7) M) of losartan (an AT(1)-receptor antagonist) or PD123319 (an AT(2)-receptor antagonist) gradually reverts inhibition by Ang H. Both the phospholipase C (PLC) inhibitor U-73122 (10(-6) M) and the inhibitor of protein kinase C (PKC) staurosporine (10-7 M) prevent inhibition of the Ca2+ pump by Ang H. Incubation of the previously isolated membranes with a PKC activator-the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (10(-8) M)-mimics the inhibition found with Ang H, and the effects of the compounds are not additive. Taken as a whole, these results indicate the Ang H inhibits Ca2+-ATPase by activation of a PKC system present in primed state in these membranes after binding of the hormone to losartan- and PD123319-sensitive receptors coupled to a PLC. Therefore, inhibition of the basolateral membrane Ca2+-ATPase by kinase-mediated phosphorylation appears to be one of the pathways by which Ang II promotes an increase in the cytosolic Ca2+ concentration of proximal tubule cells. (C) 2004 Published by Elsevier B.V.