Characterization of the receptor for glucagon-like peptide-1(7-36)amide on plasma membranes from rat insulinoma-derived cells by covalent cross-linking.
125I-Labelled glucagon-like peptide-1(7-36)amide was cross-linked to a specific binding protein in plasma membranes prepared from RINm5F rat insulinoma-derived cells using disuccinimidyl suberate. Consistent with the presence of a single class of binding site on the surface of intact cells, only a single radiolabelled band at Mr63,000 was identified by SDS-PAGE after solubilization of the ligand-binding protein complex. The band was not observed when 10nM glucagon-like peptide-1(7-36)amide was included in the binding assay, but 1 microM concentrations of glucagon-like peptide-1(1-36)amide, glucagon-like peptide-2 and glucagon did not decrease the intensity of labelling. No change in the mobility of the band was observed under reducing conditions, suggesting that the binding protein in the receptor is not attached to other subunits via disulphide bonds. In control incubations using plasma membranes from pig intestinal epithelial cells, which do not contain specific binding sites for glucagon-like peptide-1(7-36)amide, no cross-linked ligand-binding protein complex was observed.
Göke
R
R
Clinical Research Group for Gastrointestinal Endocrinology, Max-Planck-Gesellschaft, University of Göttingen, Federal Republic of Germany.
Cole
T
T
Conlon
J M
JM
D007516:Adenoma, Islet Cell / Q000473:pathology*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D003432:Cross-Linking Reagents; D005934:Glucagon; D052216:Glucagon-Like Peptide 1; D000067757:Glucagon-Like Peptide-1 Receptor; D004763:Glucagon-Like Peptides; D007340:Insulinoma / Q000032:analysis / Q000473:pathology* / Q000648:ultrastructure; D010455:Peptides / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000032:analysis / Q000378:metabolism; D018027:Receptors, Glucagon
D003432:Cross-Linking Reagents; C000593835:Glp1r protein, rat; D000067757:Glucagon-Like Peptide-1 Receptor; D010455:Peptides; D011956:Receptors, Cell Surface; D018027:Receptors, Glucagon; D004763:Glucagon-Like Peptides; D052216:Glucagon-Like Peptide 1; D005934:Glucagon; C055454:glucagon-like peptide 1 (1-36)amide