Selective proton permeability and pH regulation of the influenza virus M2 channel expressed in mouse erythroleukaemia cells.
1. The M2 protein of influenza A virus is implicated in transmembrane pH regulation during infection. Whole-cell patch clamp of mouse erythroleukaemia cells expressing the M2 protein in the surface membrane showed a conductance due to M2 which was specifically blocked by the anti-influenza drug rimantadine. 2. The ion selectivity of the rimantadine-sensitive current through M2 was determined. Reversal potentials were close to equilibrium potentials for transmembrane pH gradients and not to those for Na+, K+ or Cl- concentration gradients. M2 permeability to Na+ relative to H+ was estimated to be less than 6 x 10(-7). 3. The M2 conductance increased as external pH decreased below 8.5 and approached saturation at an external pH of 4, effects attributable to increased permeability due to increased driving potential and to activation by low external pH. Both activation and permeation could be described by interaction of protons with sites on M2, with apparent dissociation constants of approximately 0.1 microM and 1 microM, respectively, under physiological conditions. 4. The M2 protein can transfer protons selectively across membranes with the H+ electrochemical gradient, properties consistent with its role in modifying virion and trans-Golgi pH during virus infection.
Chizhmakov
I V
IV
National Institute for Medical Research, London, UK.
Geraghty
F M
FM
Ogden
D C
DC
Hayhurst
A
A
Antoniou
M
M
Hay
A J
AJ
D000818:Animals; D002463:Cell Membrane Permeability; D004553:Electric Conductivity; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000502:physiology*; D007473:Ion Channels / Q000502:physiology; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D018408:Patch-Clamp Techniques; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000187:drug effects; D012299:Rimantadine / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000502:physiology*; D000086082:Viroporin Proteins
D007473:Ion Channels; D011994:Recombinant Proteins; D012299:Rimantadine; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins