Isolation of matrix protein M1 from influenza viruses by acid-dependent extraction with nonionic detergent.
Influenza viruses were disrupted layer by layer with the nonionic detergent NP-40 at fixed pH. Treatment of the virions with NP-40 at neutral or mildly alkaline pH (6.8-8.0) yielded viral core structures containing M1 protein. The matrix M1 protein was selectively extracted from cores at acidic pH 3.0-4.5 with citrate, acetate, and phosphate buffers or with morpholinoethanesulfonic acid. The resulting M1 protein sedimented in a glycerol gradient with a coefficient of 2.8 S and most likely existed as a monomeric form of the 27,000-Da polypeptide. An antigenic map of the monomeric protein M1 tested with a panel of monoclonal anti-M1 antibodies was found to be similar to those of the assembled M1 protein in whole virions. The isolated M1 protein retained biological properties and inhibited the RNA polymerase activity of viral RNP. This transcription-inhibition function of M1 monomers was specifically restricted by one of the monoclonal antibodies studied.
Zhirnov
O P
OP
D.I. Ivanovsky Virology Institute, Moscow, USSR.
D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D002499:Centrifugation, Density Gradient; D003902:Detergents; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000737:chemistry* / Q000235:genetics; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000737:chemistry; D012995:Solubility; D014158:Transcription, Genetic; D014763:Viral Matrix Proteins / Q000276:immunology / Q000302:isolation & purification*; D014771:Virion / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D003902:Detergents; D014763:Viral Matrix Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40