Characterization of influenza virus neuraminidases: peptide changes associated with antigenic divergence between early and late N2 neuraminidases.
Neuraminidases (EC 3.2.1.18) of 1957, 1960, and 1969 influenza virus strains were isolated after proteolytic digestion of viral hemagglutinin. Each neuraminidase was recovered with a final yield of about 15% and had similar specific activities. Immunization of rabbits with the neuraminidases elicited monospecific neuraminidase antibodies, with no antibodies to viral hemagglutinin. Further evidence of purity was the existence of only a single component, about 50,000 daltons in size, when reduced neuraminidase preparations were examined by sodium dodecyl sulfate acrylamide gel electrophoresis. However, storage of neuraminidase in solution resulted in the appearance of slightly smaller degradation products. Preparations of each neuraminidase were denatured under reducing conditions, and exposed sulfhydryl residues were blocked by reaction with (14)C-iodoacetamide. After tryptic digestion, peptide maps were prepared for the neuraminidases, and the (14)C-labeled cysteinyl peptides were then identified by autoradiography. About 20 peptides were present, in agreement with the number predicted from amino acid analysis for neuraminidase subunits of only one type. The 1957 and 1960 neuraminidases exhibited a small antigenic divergence from each other, and maps of their cysteinyl peptides appeared to be identical. The 1969 neuraminidase exhibited considerable antigenic divergence from the other two neuraminidases, and maps of 1969 neuraminidase peptides revealed two major and several minor differences from the other maps. Thus, antigenic divergence between the neuraminidases of Asian and Hong Kong influenza viruses is associated with a small number of changes in the primary structure of the neuraminidase subunit.
D000956:Antigens, Viral; D001345:Autoradiography; D002250:Carbon Radioisotopes; D002499:Centrifugation, Density Gradient; D002845:Chromatography; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D006385:Hemagglutination Inhibition Tests; D008970:Molecular Weight; D009439:Neuraminidase / Q000032:analysis* / Q000302:isolation & purification; D009975:Orthomyxoviridae / Q000201:enzymology* / Q000276:immunology; D010455:Peptides / Q000032:analysis*; D014316:Tritium
D000956:Antigens, Viral; D002250:Carbon Radioisotopes; D010455:Peptides; D014316:Tritium; D009439:Neuraminidase