TY - JOUR
T1 - Characterization of the receptor for glucagon-like peptide-1(7-36)amide on plasma membranes from rat insulinoma-derived cells by covalent cross-linking
AU - Goke, R.
AU - Cole, T.
AU - Conlon, J. M.
PY - 1989
Y1 - 1989
N2 - 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 M(r) 63,000 was identified by SDS-PAGE after solubilization of the ligand-binding protein complex. The band was not observed when 10 nM glucagon-like peptide-1(7-36)amide was included in the binding assay, but 1 μM 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.
AB - 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 M(r) 63,000 was identified by SDS-PAGE after solubilization of the ligand-binding protein complex. The band was not observed when 10 nM glucagon-like peptide-1(7-36)amide was included in the binding assay, but 1 μM 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.
UR - http://www.scopus.com/inward/record.url?scp=0024600552&partnerID=8YFLogxK
U2 - 10.1677/jme.0.0020093
DO - 10.1677/jme.0.0020093
M3 - Article
C2 - 2550026
AN - SCOPUS:0024600552
SN - 0952-5041
VL - 2
SP - 93
EP - 98
JO - Journal of Molecular Endocrinology
JF - Journal of Molecular Endocrinology
IS - 2
ER -