Solubilization and characterization of CCK receptors from mouse pancreas.
To study the characteristics of the CCK receptor, plasma membranes were prepared from mouse pancreatic acini, and CCK receptors solubilized with 1% digitonin. To measure hormone binding, the solubilized receptors were incubated with 125I-CCK at 4 degrees C and the hormone-receptor complex was precipitated with 10% polyethylene glycol. Specific 125I-CCK binding by the solubilized CCK receptor was compared to that by the plasma membrane-bound CCK receptor. Both the solubilized and the membrane-bound receptor displayed optimal binding at an acidic pH (between 6.0 and 7.0) and showed a similar sensitivity to monovalent and divalent cations. The solubilized receptors preserved their relative specificity for CCK molecules: CCK-8 greater than CCK-33 greater than desulfated CCK-8 greater than CCK-4. However, the soluble CCK receptor had a lower binding affinity than plasma membrane-bound receptor. Solubilized receptors preserved their relative specificity for inhibitors of CCK binding and action: dibutyryl cyclic GMP greater than N-CBZ-tryptophan greater than proglumide. Solubilized receptors had affinities for these antagonists that were similar to receptors on intact plasma membranes. These data indicate, therefore, that the specific binding properties of the CCK receptor are inherent to the solubilized glycoprotein molecules.
Szecòwka
J
J
Goldfine
I D
ID
Williams
J A
JA
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D002766:Cholecystokinin / Q000037:antagonists & inhibitors; D003902:Detergents; D006023:Glycoproteins / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D008297:Male; D051379:Mice; D010179:Pancreas / Q000032:analysis*; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000302:isolation & purification*; D011949:Receptors, Cholecystokinin; D012995:Solubility; D013552:Swine; D013997:Time Factors
D003902:Detergents; D006023:Glycoproteins; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D011949:Receptors, Cholecystokinin; D002766:Cholecystokinin