Characterization of Rauscher murine leukemia virus envelope glycoprotein receptor in membranes from murine fibroblasts

J Virol. 1978 Dec;28(3):686-96. doi: 10.1128/JVI.28.3.686-696.1978.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Calcium / pharmacology
  • Cell Line
  • Detergents / pharmacology
  • Glycoproteins / metabolism*
  • Glycoside Hydrolases / pharmacology
  • Hydrogen-Ion Concentration
  • Kinetics
  • Peptide Hydrolases / pharmacology
  • Phospholipases / pharmacology
  • Rauscher Virus*
  • Receptors, Virus / metabolism*
  • Temperature
  • Viral Proteins / metabolism*

Substances

  • Detergents
  • Glycoproteins
  • Receptors, Virus
  • Viral Proteins
  • Phospholipases
  • Glycoside Hydrolases
  • Peptide Hydrolases
  • Calcium