Identification of the gB homologues of equine herpesvirus types 1 and 4 as disulphide-linked heterodimers and their characterization using monoclonal antibodies.
Equine herpesvirus types 1 and 4 (EHV-1 and EHV-4) labelled with [14C]glucosamine were purified from infected cell culture medium and profiles of their structural proteins were obtained that enabled identification of the major glycoproteins. Nine glycosylated polypeptides were identified for each virus. Preparations of the purified viruses each contained a glycoprotein which was linked by disulphide bonds, as determined by diagonal gel electrophoresis under reducing/non-reducing conditions. High Mr forms of this glycoprotein were detected for EHV-1 when the sample was not heated. The EHV-1 protein consisted of three polypeptides of Mr 108K, 76K and 58K and the EHV-4 protein consisted of three polypeptides of Mr 112K, 74K and 61K. Western blotting and immunoprecipitation with monoclonal antibodies confirmed that the EHV-1 gB homologue migrates with an apparent Mr of 108K (140K under non-reducing conditions) but is cleaved to give glycoproteins of 76K and 58K which are held together by disulphide bonds. The EHV-4 gB homologue consists of a 112K glycoprotein which is cleaved to give glycoproteins of 74K and 61K which are also linked by disulphide bonds.
Meredith
D M
DM
Department of Microbiology, University of Leeds, U.K.
Stocks
J M
JM
Whittaker
G R
GR
Halliburton
I W
IW
Snowden
B W
BW
Killington
R A
RA
D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D004220:Disulfides / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology; D006564:Herpesviridae / Q000032:analysis*; D004861:Herpesvirus 1, Equid / Q000032:analysis* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests; D014764:Viral Proteins / Q000032:analysis* / Q000276:immunology; D015678:Viral Structural Proteins
D000911:Antibodies, Monoclonal; D004220:Disulfides; D006023:Glycoproteins; D014764:Viral Proteins; D015678:Viral Structural Proteins