Identification of feline herpesvirus type 1-hemagglutinin.
The crude hemagglutinin of feline herpesvirus type 1 (FHV-1), solubilized from infected fcwf-4 cells by detergents, was partially purified by three kinds of chromatographic methods. Lectin-affinity chromatography showed the hemagglutination (HA) activity in fractions, which was bound to Concanavalin A-sepharose and then eluted by alpha-methyl D-mannoside, suggesting that the hemagglutinin might include a glycoprotein. Ion-exchange and gel-exclusion chromatographies were also capable of purifying the detergent-soluble crude hemagglutinin. When peak HA fractions, which were obtained from each of the three procedures, were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the gel-exclusion chromatography was the most effective method. Electrophoreic analysis also showed only one band of 59,000 (59K) molecular weight protein, which was commonly observed in the three partially purified hemagglutinins with silver staining. In addition, the 59K protein band was clearly recognized in immunoblot analysis of the infected cell lysates using infected cat serum. These observations suggest that the FHV-1 detergent-soluble hemagglutinin from infected fcwf-4 cells may be closely related to a 59K immunogenic glycoprotein.
Horimoto
T
T
Kasaoka
T
T
Tuchiya
K
K
Takahashi
E
E
D000818:Animals; D002415:Cats; D002478:Cells, Cultured; D002845:Chromatography; D002850:Chromatography, Gel; D003902:Detergents; D006389:Hemagglutinins, Viral / Q000302:isolation & purification*; D006564:Herpesviridae / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000302:isolation & purification*; D008970:Molecular Weight; D012995:Solubility
D003902:Detergents; D006389:Hemagglutinins, Viral; D008562:Membrane Glycoproteins