Characterization of cholecystokinin receptor subunits on pancreatic plasma membranes.
To determine the size and subunit structure of the pancreatic cholecystokinin (CCK) receptor, 125I-CCK33 was covalently cross-linked to its receptor on mouse pancreatic acinar plasma membranes utilizing the bifunctional cross-linker disuccinimidyl suberate. When CCK was cross-linked at pH 7.4 to either purified plasma membranes or to isolated pancreatic acini and then followed by preparation of plasma membranes, the major labeled protein band revealed by polyacrylamide gel electrophoresis was Mr = 120,000 in the absence of reducing agent and Mr = 80,000 in the presence of reducing agent. A similar banding pattern was also observed when different cross-linkers, ethylene glycol bis(succinimidyl succinate) or dithiobis (succinimidyl propionate), were employed. At pH 6.0, where CCK binding to its receptors is optimal, the labeling pattern was similar to that seen at 7.4, although the two bands were more heavily labeled. Both the binding of CCK to its receptors on plasma membranes and the appearance of the two cross-linked proteins on gels were inhibited in a parallel manner by increasing concentrations of unlabeled CCK8; similar results were observed with dibutyryl cyclic GMP, a competitive inhibitor of CCK binding and action. The data indicate, therefore, that the CCK receptor possesses subunit structure whereby an Mr = 76,000 binding subunit is linked to an Mr = 40,000 nonbinding subunit by a disulfide bond.
D000818:Animals; D001667:Binding, Competitive; D002462:Cell Membrane / Q000378:metabolism; D002766:Cholecystokinin / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008297:Male; D051379:Mice; D008970:Molecular Weight; D010179:Pancreas / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D011949:Receptors, Cholecystokinin
D046911:Macromolecular Substances; D011956:Receptors, Cell Surface; D011949:Receptors, Cholecystokinin; D002766:Cholecystokinin; D004229:Dithiothreitol