Adrenal cortex adenylate cyclase: solubilization of adenylate cyclase and guanyl nucleotide binding sites

Abstract

Adenylate cyclase activity and binding sites for 5'-guanylyl-imidiphosphate [Cpp(NH)p] have been solubilized from bovine adrenal cortex membranes with Triton N-101. Solubilized adenylate cyclase is stimulated by Gpp(nh)p and fluoride ion but not by ACTH. Pretreatment of particulate membrane fractions with Gpp(NH)p, hormone and Gpp(NH)p or fluoride prior to detergent extraction leads to an activated solubilized enzyme which is nearly independent from added stimulants. Adenylate cyclase solubilized from membranes pretreated with Gpp(NH)p can be separated from guanyl nucleotide binding sites without loss of activity.