title: Consolidation of intramolecular disulfide bonds in influenza virus hemagglutinin as an element of intracellular maturation.
abstract: Electrophoretic behavior of influenza virus hemagglutinin during SDS electrophoresis in polyacrylamide gel is critically dependent on the life time in the infected cells and also on the conditions of sample preparation and analysis. During electrophoresis of total cell lysate proteins under nonreducing conditions the short-labeled hemagglutinin is detected as multiple bands, electrophoretic mobility of most of them being lower than that of hemagglutinin of viral particles. This heterogeneity failed to be detected during electrophoresis under reducing conditions which is indicative of the differences in the number or direction of intramolecular disulfide bonds between short-labeled and mature hemagglutinin molecules. After chasing at 37 or 20 degrees hemagglutinin gradually assumes an electrophoretic character identical to that of virion protein. Chasing at 0 degrees or the substitution of parafluorophenyl alanine for phenylalanine in the maintenance medium during labeling prevents maturation. At the same time, both iodacetamide perfusion of infected cells and the preparation of nuclei-free extract prior to SDS lysis result in a marked increase in the yield of disulfide mature short-labeled hemagglutinin. These results suggest that disulfide maturation in hemagglutinin proceeds in two stages: a relatively rapid (with respect to synthesis completion) formation of intramolecular disulfide bonds as such followed by a much slower consolidation of bridges against the action of endogenous cell reductants which activate during lysis. Consolidation may be caused by two factors: trimerization of hemagglutinin monomers or their covalent post-translational modifications.
authors.affiliation: D.I. Ivanovsky Institute of Virology, USSR Academy of Medical Sciences, Moscow.
authors.forename: L M
authors.initials: LM
authors.lastname: Selimova
authors.forename: A A
authors.initials: AA
authors.lastname: Tashenova
authors.forename: V M
authors.initials: VM
authors.lastname: Zaides
mesh_terms: D000818:Animals; D002642:Chick Embryo; D004220:Disulfides / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000302:isolation & purification; D009980:Influenza A virus / Q000502:physiology*; D008970:Molecular Weight; D011498:Protein Precursors / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D004220:Disulfides; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D011498:Protein Precursors; D014759:Viral Envelope Proteins