Similar characteristics of guanine nucleotide regulatory sites involved in adenylate cyclase activation, specific GTPase activity, and cholecystokinin binding in rat pancreatic plasma membranes.

The specificity of guanine nucleotide regulatory site(s) of rat pancreatic plasma membranes involved in adenylate cyclase activation, basal and cholecystokinin (CCK)-stimulated specific GTPase activity, and [125I]-CCK-33 binding was documented. The Km (for GTP) and Ki (for other nucleotides) of basal and CCK-8-dependent GTPase showed similar specificity, decreasing in the order GTP gamma S approximately GTP approximately Gpp[NH]p greater than ITP greater than GDP beta S greater than UTP, suggesting the identity of basal and CCK-stimulated GTPase activities. The same potency order for these nucleotides was obtained when tested as activator (Ka) or inhibitor (Ki for GDP beta S) of adenylate cyclase, in the presence of CCK-8. The IC 50 of these nucleotides on the binding of [125I]-CCK-33 indicated a similar specificity for nucleotide binding sites interacting with CCK receptors (GTP gamma S approximately Gpp[NH]p approximately GTP greater than ITP approximately GDP beta S greater than UTP). In membranes preactivated with 0.3 micron CCK-8 and 30 micron Gpp[NH]p or GTP gamma S, then washed free of hormone and unbound nucleotide, persistent effects of the nonhydrolyzable nucleotides were observed on the three activities tested. The present data indicate that the guanine nucleotide regulatory units involved in adenylate cyclase activation, GTPase activity and CCK binding have similar properties in rat pancreatic plasma membranes.