Analysis of a glycoprotein of human herpesvirus 6 (HHV-6) using monoclonal antibodies.
The virus-specific polypeptides of human herpesvirus 6 (HHV-6) were studied. Six hybridoma clones secreting monoclonal antibodies were established. Localized cytoplasmic antigen and surface antigens were found in and on the infected cells, respectively, by using the immunofluorescent test with those antibodies. In the neutralization test, two clones (OHV3 and OHV9) neutralized HHV-6, but the other five monoclonal antibodies did not. When infected cells were labeled with [35S]methionine and the antigens were immunoprecipitated by OHV3 and OHV9 followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, two polypeptides with molecular weights of 98,000 (98K) and 92K were detected. Furthermore, it was found that these polypeptides were glycosylated, because they were labeled with [14C]mannose. Pulse-chase studies revealed that precursor molecules were cotranslationaly glycosylated to produce 98K glycoprotein and they were replaced by 92K glycoprotein. Endo-beta-N-acetylglucosaminidases H and F reduced those glycoproteins to putative precursor molecules of 75 kDa. This indicates that N-linked high-mannose sugars associate to 98K and 92K glycoproteins.
Okuno
T
T
Department of Virology, Research Institute for Microbial Diseases, Osaka University, Japan.
Sao
H
H
Asada
H
H
Shiraki
K
K
Takahashi
M
M
Yamanishi
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D005312:Fetal Blood / Q000166:cytology; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology; D015654:Herpesvirus 6, Human / Q000032:analysis* / Q000276:immunology; D006801:Humans; D006825:Hybridomas; D008214:Lymphocytes / Q000382:microbiology; D051379:Mice; D009500:Neutralization Tests; D011233:Precipitin Tests; D014764:Viral Proteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D014764:Viral Proteins