Demonstration of three major species of pseudorabies virus glycoproteins and identification of a disulfide-linked glycoprotein complex.
The glycoproteins of pseudorabies virus (PRV) Phylaxia were characterized with monoclonal antibodies as specific reagents. Three major structural glycoproteins with molecular weights of 155,000 (155K) (gC), 122K (gA), and 90K (gB) could be identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions. We investigated the processing of glycoproteins gA, gB, and gC by in vitro translation, pulse-chase experiments, and in the presence of the ionophore monensin which inhibits glycosylation. gA and gB were found to compose a single polypeptide, whereas gC was found to be a disulfide-linked glycoprotein complex. Immunoprecipitates formed with the aid of anti-gC monoclonal antibodies gave rise to three glycoprotein bands (gC0 [120K], gC1 [67K], and gC2 [58K]) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Limited proteolysis of gC0, gC1, and gC2 resulted in peptide maps of gC0 related to those of both gC1 and gC2. No common peptide bands between gC1 and gC2, however, were seen. We suggest that (i) gC1 and gC2 arise by proteolytic cleavage from the same precursor molecule and stay joined via disulfide bridges and (ii) gC0 is an uncleaved precursor.
Lukàcs
N
N
Thiel
H J
HJ
Mettenleiter
T C
TC
Rziha
H J
HJ
D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D002460:Cell Line; D004220:Disulfides / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis*; D011558:Herpesvirus 1, Suid / Q000032:analysis* / Q000235:genetics; D007668:Kidney; D046911:Macromolecular Substances; D008970:Molecular Weight; D014176:Protein Biosynthesis; D051381:Rats; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics; D015678:Viral Structural Proteins
D000911:Antibodies, Monoclonal; D004220:Disulfides; D000939:Epitopes; D006023:Glycoproteins; D046911:Macromolecular Substances; D014759:Viral Envelope Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C042966:pseudorabies virus glycoproteins