Adrenal cortex adenylate cyclase. In vitro modification of the enzyme by cholera toxin

Pretreatment of rat adrenal particulate fractions with cholera toxin in vitro markedly changed the properties of the membrane-bound adenylate cyclase. The basal activity of the enzyme was increased after cholera toxin treatment. The main action of the toxin was on the Vmax of the enzyme. In the absence of added GTP Lineweaver-Burk plots indicate a deviation from normal Michaelis-Menten kinetics with respect to substrate, the slopes being concave downward for control and toxin-treated membranes. Although hormonal stimulation of the enzyme was diminished in toxin-treated membranes, the hormone receptors were still functionally active as revealed after addition of Gpp(NH)p, GTP or GTPgammaS. The response to NaF was decreased in the toxin-treated membranes. Whereas GTP behaves as an antagonist (or a partial agonist with low intrinsic activity) compared to Gpp(NH)p in control membranes, it has similar intrinsic activity as Gpp(NH)p in the toxin-treated membranes. The results indicate that cholera toxin modification of the adenylate cyclase complex is located at the guanyl nucleotide sites or factors controlling the turnover of GTP at these sites. Cholera toxin modification may be a useful tool to investigate the role of guanyl nucleotide sites in the regulation of adenylate cyclase activity.