Reconstitution of the influenza virus M2 ion channel in lipid bilayers.
M2, an integral membrane protein of influenza A virus, was purified from either influenza A virus-infected CV-1 cells or from Spodoptera frugiperda (Sf9) cells infected with a recombinant-M2 baculovirus. The purified protein, when incorporated into phospholipid bilayer membranes, produced ion-permeable channels with the following characteristics: (1) The channels appeared in bursts during which unit conductances of diverse magnitudes (25-500 pS) were observed. (2) The most probable open state was usually the lowest unit conductance (25-90 pS). (3) The channels were selective for cations; tNa = 0.75 when 150 mM NaCl bathed both sides of the membrane. (4) Amantadine reduced the probablity of opening of the high conductance state and also the conductance of the most probable state. (5) Reducing pH increased the mean current through the open channel as well as the conductance of the most probable state. (6) The sequence of selectivity for group IA monovalent cations was Rb > K > Cs approximately Na > Li. The pH activation, amantadine block and ion selectivity of the M2 protein ion channel in bilayers are consistent with those observed on expression of the M2 protein in oocytes of Xenopus laevis as well as for those predicted for the proposed role of an ion channel in the uncoating process of influenza virus. The finding that the M2 protein has intrinsic ion channel activity supports the hypothesis that it has ion channel activity in the influenza virus particle.
Tosteson
M T
MT
Laboratory for Membrane Transport, Harvard Medical School, Boston, Massachusetts 02115.
Pinto
L H
LH
Holsinger
L J
LJ
Lamb
R A
RA
D000547:Amantadine / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D002414:Cations, Monovalent / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009980:Influenza A virus / Q000737:chemistry*; D007473:Ion Channels / Q000378:metabolism*; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D018411:Spodoptera; D013329:Structure-Activity Relationship; D014763:Viral Matrix Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000086082:Viroporin Proteins
D000547:Amantadine; D002414:Cations, Monovalent; D007473:Ion Channels; D008051:Lipid Bilayers; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins