Purification of the 110-kilodalton glycoprotein receptor for mouse hepatitis virus (MHV)-A59 from mouse liver and identification of a nonfunctional, homologous protein in MHV-resistant SJL/J mice.
The receptor for mouse hepatitis virus strain A59 (MHV-A59) is a 110- to 120-kilodalton (kDa) glycoprotein which is expressed in MHV-susceptible mouse strains on the membranes of hepatocytes, intestinal epithelial cells, and macrophages. SJL/J mice, which are highly resistant to MHV-A59, were previously shown to lack detectable levels of receptor by using either solid-phase virus receptor assays or binding of a monoclonal anti-receptor antibody (MAb) which blocks infection of MHV-susceptible mouse cells. This MAb was used for affinity purification of the receptor glycoprotein from livers of MHV-susceptible Swiss Webster mice. The MHV receptor and an antigenically related protein of 48 to 58 kDa were copurified and then separated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The first 15 amino acids of the receptor were sequenced, and a synthetic peptide of this amino acid sequence was prepared. Rabbit antiserum made against this peptide bound to the MHV receptor glycoprotein and the 48- to 58-kDa protein from livers of MHV-susceptible BALB/c mice and Swiss Webster mice and from the intestinal brush border of BALB/c mice. In immunoblots of intestinal brush border and hepatocyte membranes of MHV-resistant SJL/J mice, the antibody against the amino terminus of the receptor identified proteins that are 5 to 10 kDa smaller than the MHV receptor and the 48- to 58-kDa related protein from Swiss Webster or BALB/c mice. Thus, SJL/J mice express a protein which shares some sequence homology with the MHV receptor but which lacks virus-binding activity and is not recognized by the blocking anti-receptor MAb. These results suggest that resistance of SJL/J mice to MHV-A59 may be due to absence or mutation of the virus-binding domain in the nonfunctional receptor homolog in SJL/J mice.
Williams
R K
RK
Department of Pathology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814.
Jiang
G S
GS
Snyder
S W
SW
Frana
M F
MF
Holmes
K V
KV
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D004198:Disease Susceptibility; D005260:Female; D006524:Hepatitis, Viral, Animal / Q000276:immunology*; D007422:Intestines / Q000276:immunology; D008099:Liver / Q000382:microbiology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D008815:Mice, Inbred Strains; D008871:Microvilli / Q000276:immunology; D008970:Molecular Weight; D006517:Murine hepatitis virus / Q000502:physiology*; D011991:Receptors, Virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D008562:Membrane Glycoproteins; D011991:Receptors, Virus