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

Arch Virol. 2008;153(10):1977-80. doi: 10.1007/s00705-008-0214-7. Epub 2008 Sep 30.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cold Temperature
  • Detergents / pharmacology
  • Hemagglutinin Glycoproteins, Influenza Virus / isolation & purification*
  • Influenza A virus / chemistry*
  • Influenza A virus / ultrastructure
  • Macromolecular Substances / isolation & purification
  • Membrane Lipids / isolation & purification
  • Microscopy, Electron, Transmission
  • Viral Matrix Proteins / isolation & purification*
  • Virion / chemistry*
  • Virion / ultrastructure

Substances

  • Detergents
  • Hemagglutinin Glycoproteins, Influenza Virus
  • M1 protein, Influenza A virus
  • Macromolecular Substances
  • Membrane Lipids
  • Viral Matrix Proteins
  • hemagglutinin, human influenza A virus
  • viral envelope lipids