Abstract
Inhibitory effect of guanyl nucleotides toward adenylate cyclase activity of Chinese hamster ovary cell membranes activated in vitro by cholera toxin.
Highlights
The adenylate cyclase activity of crude membranes prepared from Chinese Hamster Ovary (CHO) cells in culture is enhanced about a-fold with 0.1 mM GTP and
We present evidence that incubation of isolated CHO membranes under conditions which result in maximal cholera toxin activation converts the adenylate cyclase system to a form which is sensitive to inhibition by GTP and
In vitro cholera toxin treatment of rat liver homogenates [22], which presumably contain sufficient endogenous GTP, results in an activated adenylate cyclase which exhibits an inhibitory response to Gpp(NH)p
Summary
Cholera toxin has been shown to activate the adenylate cyclase system of a number of eukaryotic cells (see Ref. 8). Activation of adenylate cyclase by cholera toxin can be provoked by exposure of intact cells to choleragen or by toxin treatment of broken cell preparation when performed in the presence of NAD’. In this communication, we report a potent system for the cell-free activation of adenylate cyclase activity by cholera toxin with membranes prepared from Chinese Hamster Ovary cells growing in culture. Of particular interest is the finding that following cell-free activation by cholera toxin, the guanyl nucleotides GTP and Gpp(NH)p’ both exert an inhibitory effect on adenylate cyclase activity
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