Characterization of vaccinia virus glycoproteins by monoclonal antibody precipitation.
Monoclonal antibodies were used to characterize vaccinia virus glycoproteins known to be incorporated into the envelope of extracellular enveloped vaccinia (EEV) virus. The 89K hemagglutinin, 42K, and 23-28K glycoproteins were predominantly expressed as late vaccinia proteins. The 89K glycoprotein was sulfated and phosphorylated but not acylated. 89K precursor proteins of 32K, 41.5K, and 52K were detected. The former had a molecular weight expected from the deduced amino acid sequence of the hemagglutinin gene. A 76K glycoprotein that did not contain methionine and was not sulfated or phosphorylated was precipitated late in infection by the anti-hemagglutinin monoclonal antibody. The appearance of this protein was inhibited by rifampicin and it may thus result from 89K cleavage. A 220K complex contained some or all of the hemagglutinin gene products linked by disulfide bonds. The 42K glycoprotein was not sulfated or phosphorylated but was acylated. This glycoprotein was disulfide bonded with the EEV 37K nonglycosylated envelope protein. The 23-28K glycoprotein was not sulfated but was both phosphorylated and acylated. The 23-28K glycoprotein group of five proteins had a common protein backbone that was differentially glycosylated. Pulse-chase, glycosylation inhibition with tunicamycin, and glycosidase experiments established that the precursor to the 23-28K glycoproteins was a 21K protein. Members of this protein family formed dimers of approximately 55K through disulfide bonds.
Payne
L G
LG
Department of Virology, Karolinska Institute, School of Medicine, Stockholm, Sweden.
D000107:Acetylation; D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D006023:Glycoproteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006031:Glycosylation; D006389:Hemagglutinins, Viral / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D006825:Hybridomas; D008970:Molecular Weight; D008985:Monensin / Q000494:pharmacology; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D010766:Phosphorylation; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational; D014415:Tunicamycin / Q000494:pharmacology; D014616:Vaccinia virus / Q000276:immunology / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D014759:Viral Envelope Proteins; D014415:Tunicamycin; D008985:Monensin; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase