Characterization of bovine rotavirus VP6 and VP7 as glycoproteins using monoclonal antibodies.
Bovine rotavirus proteins were analysed by a panel of monoclonal antibodies. Glycosylated epitopes were identified on both inner and outer capsid proteins (VP6 and VP7 respectively). VP7 possessed a periodate insensitive epitope which was, however, sensitive to endoglycosidase H, mixed glycosidases and to protease treatment. This epitope was not detected on viruses grown in the presence of 2-deoxy-D-glucose or tunicamycin. An epitope was detected on VP6 which was sensitive to periodate oxidation. The blotted protein reacted with a glycan assay kit; yet the epitope was not affected by endoglycosidase H and was found on viruses grown in the presence of 2-deoxy-D-glucose or tunicamycin. These results suggest that VP7 and VP6 epitopes are carbohydrate dependent. The VP7 epitope contains an N-linked carbohydrate moiety in contrast to the VP6 epitope which appears to contain O-linked glycosyl units.
Cornaglia
E
E
Virology Section, Faculty of Veterinary Medicine, University of Montreal, Quebec, Canada.
Elazhary
Y
Y
Talbot
B
B
D000118:Acetylglucosaminidase / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D002213:Capsid / Q000737:chemistry / Q000276:immunology*; D036022:Capsid Proteins; D002417:Cattle; D003847:Deoxyglucose / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006023:Glycoproteins / Q000276:immunology; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000378:metabolism; D011233:Precipitin Tests; D012401:Rotavirus / Q000276:immunology*; D014415:Tunicamycin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D036022:Capsid Proteins; D000939:Epitopes; D006023:Glycoproteins; C061332:VP6 protein, Rotavirus; C050779:VP7 protein, Rotavirus; D010504:Periodic Acid; D014415:Tunicamycin; D003847:Deoxyglucose; C009288:metaperiodate; D006026:Glycoside Hydrolases; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D010450:Endopeptidases