Structure of bromelain-released influenza virus haemagglutinin as revealed by electrophoresis, sedimentation and electron microscopy.
Sigma bromelain (EC 3.4.22.4) was used to isolate the haemagglutinin (HA) from the MRC-11 (H3N2) and A/U.S.S.R./90/77 (H1N1) influenza A virus strains. Sedimentation analysis of bromelain-solubilized preparations revealed 9.5S and 5.5S protein components, the former being identified as the bromelain-released haemagglutinin (BHA). No residual neuraminidase (NA) activity was detected in the BHA isolated from the MRC-11 strain whereas up to 80 per cent of the enzymatically active NA was found to be preserved in the electrophoretically pure BHA isolated from the A/U.S.S.R./90/77 strain. Increased electrophoretic mobilities were exhibited by both the light and heavy chains of the BHA subunit. The difference observed in the molecular weights of the polypeptide fragments removed by bromelain from the light chains is interpreted in terms of the different depth of penetration of antigenically distinct HAs through the influenza virus lipid membrane. Splitting off of approximately 15 and 26 per cent of the sugars from the carbohydrate portions of the light and heavy chains respectively, was demonstrated. This suggested involvement of glycosidase impurities present in the bromelain preparation employed. The rod-shaped BHA molecules proved to be 110 +/- 5 Angstrom long and 40 +/- 5 Angstrom wide as measured by electron microscopy. It is proposed that the 45,000-molecular-weight polypeptide observed constantly in egg-grown influenza viruses is host actin.
D001963:Bromelains / Q000494:pharmacology; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D006389:Hemagglutinins, Viral / Q000032:analysis* / Q000302:isolation & purification; D009980:Influenza A virus / Q000187:drug effects / Q000276:immunology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis; D011487:Protein Conformation
D006389:Hemagglutinins, Viral; D010455:Peptides; D001963:Bromelains