Disulfide bonding in influenza virus proteins as revealed by polyacrylamide gel electrophoresis.
Disulfide bonding in the major proteins of influenza virus A, WSN strain, was studied by electrophoresis in sodium dodecyl sulfate-polyacrylamide gels under reducing and nonreducing conditions. The electrophoretic behavior of the proteins correlated with their localization in the virions and their chemical composition. The internal proteins of the viral particles, i.e. matrix and nucleoproteins, were shown to contain a relatively small number of cysteine residues. Electrophoresis under nonreducing conditions yielded multiple forms of the proteins which could be discriminated by small but readily observable, reproducible differences in their migration rates in the gel. the multiplicity of the protein forms was caused by the formation of intramolecular disulfide bonds in matrix and nucleoproteins that arose during or after solubilization in sodium dodecyl sulfate. On the other hand, we failed to detect native inter- and intramolecular linkages in matrix and nucleoproteins. External glycoproteins of the virions (HA and NA) had, in contrast to the internal ones, a higher number of cysteine residues and native disulfide bonds. At least three disulfide linkages were revealed in HA and NA in our experiments. In uncleaved HA all of the linkages were intramolecular. In NA at least one disulfide bond linked two identical polypeptides into a dimer. It was established that the reduction of the different disulfide linkages in HA and NA required different concentrations of the reducing agent.
D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003545:Cysteine / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000032:analysis; D006389:Hemagglutinins, Viral / Q000032:analysis; D009980:Influenza A virus / Q000032:analysis*; D009439:Neuraminidase / Q000032:analysis; D009698:Nucleoproteins / Q000032:analysis; D014763:Viral Matrix Proteins; D014764:Viral Proteins / Q000032:analysis
D006023:Glycoproteins; D006389:Hemagglutinins, Viral; D009698:Nucleoproteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; D009439:Neuraminidase; D003545:Cysteine