Biochemical study of the Feline Herpesvirus 1. Identification of glycoproteins by affinity

Feline Viral Rhinotracheitis herpesvirus (Feline Herpesvirus 1; FHV-1) was purified by rate zonal isopycnic centrifugation. Viral nucleocapsids were isolated from the nuclei and purified. Analysis of the purified and radiolabelled viral polypeptides and glycoproteins by gradient SDS polyacrylamide gel electrophoresis revealed that 23 viral proteins and 6 glycoproteins were present in the whole virus particle. Tween 80-diethylether treatment removed the major glycoproteins. FHV-1 viral glycoproteins were detected using [14C] glucosamine and a lectin binding procedure. The fluorescein isothiocyanate-Concanavalin A (FITC-Con A) specificity was checked by trials on other viruses whose glycoproteins had already been described. The FITC-Con A allows a very simplified procedure for glycoprotein determination after slab acrylamide gel electrophoresis and can be used prior to Coomassie blue staining of proteins.