Characterization of the structural proteins of porcine epizootic diarrhea virus, strain CV777.
Pig epizootic diarrhea virus cannot be grown in cell culture; for its characterization, intestinal perfusate material from a pig infected with the strain CV777 had to be used. In isopyknic sucrose gradients, a peak of virus-specific ELISA activity was detected at a density of 1.17 g/ml. Using immunoprecipitation of radioiodinated-purified virus material followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 3 proteins of low molecular weight (20,000 to 32,000 daltons [D]) were found; after blotting nitrocellulose and glycoprotein identification with concanavalin A and horseradish peroxidase, 1 of the proteins (23,000 D) gave a signal. Another protein of 58,000 D was encountered, which was the only protein binding an RNA probe. Finally, a protein of 85,000 D was visible, associated with minor bands of about 110,000 and 135,000 D in most experiments. Using the concanavalin A-blotting technique, the same bands were visualized. The demonstration of a polydisperse cluster of proteins from 20,000 to 32,000 D (of which at least 1 is glycosylated), of glycosylated proteins from 85,000 to 135,000 D, and of an RNA-binding protein of 58,000 D is taken as structural evidence that pig epizootic diarrhea virus should be classified with the Coronaviridae, irrespective of the apparent lack of an antigenic relationship with other members of that family.
D000818:Animals; D002352:Carrier Proteins / Q000032:analysis / Q000302:isolation & purification; D003332:Coronaviridae / Q000032:analysis* / Q000145:classification / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000032:analysis / Q000302:isolation & purification; D008970:Molecular Weight; D016601:RNA-Binding Proteins; D013552:Swine / Q000382:microbiology; D014764:Viral Proteins / Q000032:analysis* / Q000302:isolation & purification; D015678:Viral Structural Proteins
D002352:Carrier Proteins; D006023:Glycoproteins; D016601:RNA-Binding Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins