Identification of a gastrin binding protein in porcine gastric mucosal membranes by covalent cross-linking with iodinated gastrin.
A gastrin binding protein (GBP) has been identified in detergent extracts of porcine gastric mucosal membranes by covalent cross-linking to 125I-[Nle15]gastrin with disuccinimidyl suberate. The apparent molecular weight of the cross-linked complex (80,000) is uneffected by reduction suggesting that the GBP is not composed of disulfide-bonded subunits. Subtraction of the molecular weight of 125I-gastrin indicates that the molecular weight of the GBP is 78,000. A similar molecular weight has been observed previously for the gastrin receptor (74,000) on intact canine parietal cells and plasma membranes therefrom, and for the receptor for the related hormone cholecystokinin (76,000-85,000) on pancreatic acinar membranes under reducing conditions. The similarity in molecular weight between the gastrin receptor and the solubilized GBP suggests that the latter protein is probably the gastrin receptor. However, the concentration (2 microM) of [Nle15]gastrin required for 50% inhibition of cross-linking of gastrin to the GBP solubilized in 0.1% Triton X-100 is 200-fold greater than the value (10 nM) observed for the gastrin receptor on isolated canine gastric parietal cells. A lower concentration (0.3 microM) of [Nle15]gastrin was required to inhibit cross-linking in a milder detergent (0.4% digitonin, 0.08% cholate). Thus, the reduced affinity for gastrin of the putative solubilized form of the gastrin receptor appears to be a result of detergent extraction.
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004285:Dogs; D005753:Gastric Mucosa / Q000032:analysis*; D005755:Gastrins / Q000378:metabolism*; D006168:Guinea Pigs; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D011817:Rabbits; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism; D011949:Receptors, Cholecystokinin; D013552:Swine
D003432:Cross-Linking Reagents; D005755:Gastrins; D007457:Iodine Radioisotopes; D008670:Metals; D011956:Receptors, Cell Surface; D011949:Receptors, Cholecystokinin