Cold co-extraction of hemagglutinin and matrix M1 protein from influenza virus A by a combination of non-ionic detergents allows for visualization of the raft-like nature of the virus envelope.
Membrane solubilization with a mixture of cold non-ionic detergents has been applied to isolate detergent-resistant membranes from intact virus A lipid bilayer. Association of the viral envelope glycoproteins and M1 into a raft lipid-protein complex was verified via detergent insolubility experiments, and the M1:HA stoichiometry of the proposed supramolecular complex was estimated via amino acid analysis. Electron microscopy and dynamic light scattering data revealed that these lipid-protein rafts form unilamellar vesicles with HA spikes on their surfaces similar to influenza virus virions. Together, our data suggest that the cold co-extraction technique visualizes the raft-like nature of the viral envelope and demonstrates the interaction of matrix M1 protein with the envelope.
Radyukhin
V
V
Department of Chromatography, AN Belozersky Institute of Physico-Chemical Biology, Moscow State University, Leninskie Gory, Moscow, Russia. varvic@belozersky.msu.ru
Fedorova
N
N
Ksenofontov
A
A
Serebryakova
M
M
Baratova
L
L
D003080:Cold Temperature; D003902:Detergents / Q000494:pharmacology; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000302:isolation & purification*; D009980:Influenza A virus / Q000737:chemistry* / Q000648:ultrastructure; D046911:Macromolecular Substances / Q000302:isolation & purification; D008563:Membrane Lipids / Q000302:isolation & purification; D046529:Microscopy, Electron, Transmission; D014763:Viral Matrix Proteins / Q000302:isolation & purification*; D014771:Virion / Q000737:chemistry* / Q000648:ultrastructure
D003902:Detergents; D019267:Hemagglutinin Glycoproteins, Influenza Virus; C488936:M1 protein, Influenza A virus; D046911:Macromolecular Substances; D008563:Membrane Lipids; D014763:Viral Matrix Proteins; C483468:hemagglutinin, human influenza A virus; C050629:viral envelope lipids