Regulation of adenylate cyclase activity in GH1 cells by chlorpromazine and a heat-stable factor.

Abstract

An adenylate cyclase present in a PRL-producing tumor cell line, GH1, which is selectively stimulated by chlorpromazine, has been characterized with respect to several biochemical properties. The parameters studied include divalent metal ion specificity, guanyl nucleotide interaction, and sensitivity to sodium fluoride. The effects of calcium and the calcium chelator, EGTA, were also tested on the chlorpromazine response. The data reported herein establish the optimal conditions for the activation of adenylate cyclase by chlorpromazine in homogenates of GH1 cells. In addition, the results from this study identify a heat-stable protein in these cells which regulates cyclase activity. This protein, which can be released into the supernatant by the pretreatment of GH1 cells with EGTA, is an absolute requirement for chlorpromazine stimulation of adenylate cyclase activity in these cells. The activation of adenylate cyclase by chlorpromazine in homogenates of normal rat pituitaries was demonstrated as well as the presence of a protein factor which regulates this activity.