Identification and characterization of a heterogeneous population of growth hormone receptors in mouse hepatic membranes.
Covalent cross-linking of radiolabeled mouse growth hormone (125I-mGH) with the homobifunctional cross-linking agent bis(sulfosuccinimidyl)suberate (BS3) to microsomal membranes prepared from late pregnant mouse liver resulted in the labeling of three specific mGH binding proteins (receptors) with apparent Mr = 125,000, 62,000, and 56,000, as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. These same three specifically labeled proteins were present, but with slightly lower apparent molecular weights, when samples were electrophoresed in the absence of reductant. Cross-linking of 125I-mGH to plasma membrane-enriched fractions of late pregnant mouse liver resulted only in the specific labeling of the two lower molecular weight receptors. Removal of all N-linked carbohydrate with peptide: N-glycosidase F resulted in decreasing the apparent molecular weights of the three receptor forms to 110,000, 50,000, and 46,000 for the 125,000, 62,000, and 56,000 molecular weight forms of the receptor, respectively. Smaller decreases in the molecular weights of all three receptor forms were also apparent after treatment with neuraminidase. However, the differences seen in the intact forms of the growth hormone receptor were also present in the deglycosylated receptors. The relationship between the three forms of the growth hormone receptor was further investigated by comparing the fragments produced by proteolytic digestion of the cross-linked receptors with Staphylococcus aureus protease and endoproteinase Lys-C. The fragments produced from all three receptor forms had very similar molecular weights, although there were slight molecular weight differences in the fragments produced by endoproteinase Lys-C digestion. The overall similarity of the fragments produced by the proteolytic digestions suggests that the three forms of the receptor are related.
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006026:Glycoside Hydrolases / Q000378:metabolism; D007425:Intracellular Membranes / Q000032:analysis; D008099:Liver / Q000032:analysis / Q000166:cytology*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D051379:Mice; D008970:Molecular Weight; D009439:Neuraminidase / Q000378:metabolism; D011247:Pregnancy; D011986:Receptors, Somatotropin / Q000032:analysis*; D013388:Succinimides / Q000494:pharmacology
D011986:Receptors, Somatotropin; D013388:Succinimides; C035760:bis(sulfosuccinimidyl)suberate; D006026:Glycoside Hydrolases; D009439:Neuraminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase