Characterization of the detergent solubilized receptor for gastrin-releasing peptide.
Properties of detergent solubilized gastrin-releasing peptide receptor were investigated. Swiss 3T3 membranes were covalently labeled with [125I]GRP and homobifunctional cross-linkers. A major labeled protein of 75 kDa was resolved using SDS-polyacrylamide gel electrophoresis. When the same preparation was solubilized with zwitterionic detergent and analyzed under nondenaturing conditions the protein bound radioactivity was resolved in two different peaks, a major one of apparent molecular weight 220,000 (peak 1) and a minor one of 80,000 (peak 2) both containing the 75 kDa protein. Specific ligand binding activity also eluted with peak 1. These results indicate that the active form of bombesin/GRP receptor is a large complex containing the 75 kDa ligand binding domain.
Cirillo
D
D
Rorer Biotechnology, Inc., King of Prussia, PA 19406.
Naldini
L
L
Moody
T W
TW
Comoglio
P
P
Schlessinger
J
J
Kris
R
R
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D002499:Centrifugation, Density Gradient; D002793:Cholic Acids; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D003902:Detergents; D019886:Gastrin-Releasing Peptide; D007457:Iodine Radioisotopes; D009711:Nucleotides; D010455:Peptides; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D018004:Receptors, Bombesin; D017981:Receptors, Neurotransmitter / Q000032:analysis*; D012995:Solubility
D002793:Cholic Acids; D003432:Cross-Linking Reagents; D003902:Detergents; D007457:Iodine Radioisotopes; D009711:Nucleotides; D010455:Peptides; D018004:Receptors, Bombesin; D017981:Receptors, Neurotransmitter; D019886:Gastrin-Releasing Peptide; D011505:Protein-Tyrosine Kinases; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate