Characterization of Rauscher murine leukemia virus envelope glycoprotein receptor in membranes from murine fibroblasts.
Plasma membrane preparations from KA31 (mouse) cells contained receptors for the binding of Rauscher murine leukemia virus (R-MuLV) envelope glycoprotein, gp70. This binding was demonstrated by gel filtration of a mixture of the microsomal fraction of the cells and 125I-labeled gp70. A rapid and convenient assay was developed to measure the complex formation between the membrane receptors and gp70 involving specific precipitation of the complex by 3 to 4% polyethylene glycol. The complex formation was responsive to the concentrations of both the receptor and gp70 and also to changes in temperature and pH. The gp70 binding was a noncooperative, saturable process, and an association constant of 3.5 X 10(8) M-1 was estimated from the binding data. The complex formation was reversible and a near-total exchange of 125I-labeled gp70 in the complex was achieved by incubation with excess of unlabeled gp70. The complex formation was inhibited by protein denaturing agents, guanidine-hydrochloride and urea. Pretreatment of the membrane fractions with either chymotrypsin or phospholipase C led to a loss of the membrane-associated receptor activity, indicating that a lipoprotein structure was important for the receptor function, consistent with the observation that nonionic detergents strongly inhibited the complex formation.
D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D003902:Detergents / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010447:Peptide Hydrolases / Q000494:pharmacology; D010740:Phospholipases / Q000494:pharmacology; D011925:Rauscher Virus; D011991:Receptors, Virus / Q000378:metabolism*; D013696:Temperature; D014764:Viral Proteins / Q000378:metabolism*
D003902:Detergents; D006023:Glycoproteins; D011991:Receptors, Virus; D014764:Viral Proteins; D010740:Phospholipases; D006026:Glycoside Hydrolases; D010447:Peptide Hydrolases; D002118:Calcium