Solid-supported membrane technology for the investigation of the influenza A virus M2 channel activity.
Influenza A virus encodes an integral membrane protein, A/M2, that forms a pH-gated proton channel that is essential for viral replication. The A/M2 channel is a target for the anti-influenza drug amantadine, although the effectiveness of this drug has been diminished by the appearance of naturally occurring point mutations in the channel pore. Thus, there is a great need to discover novel anti-influenza therapeutics, and, since the A/M2 channel is a proven target, approaches are needed to screen for new classes of inhibitors for the A/M2 channel. Prior in-depth studies of the activity and drug sensitivity of A/M2 channels have employed labor-intensive electrophysiology techniques. In this study, we tested the validity of electrophysiological measurements with solid-supported membranes (SSM) as a less labor-intensive alternative technique for the investigation of A/M2 ion channel properties and for drug screening. By comparing the SSM-based measurements of the activity and drug sensitivity of A/M2 wild-type and mutant channels with measurements made with conventional electrophysiology methods, we show that SSM-based electrophysiology is an efficient and reliable tool for functional studies of the A/M2 channel protein and for screening compounds for inhibitory activity against the channel.
Balannik
Victoria
V
Department of Neurobiology and Physiology, Northwestern University, Hogan Hall, 2205 Tech Drive, Evanston, IL 60208-3500, USA.
Obrdlik
Petr
P
Inayat
Samsoon
S
Steensen
Catrin
C
Wang
Jun
J
Rausch
Joshua M
JM
DeGrado
William F
WF
Kelety
Bela
B
Pinto
Lawrence H
LH
D000547:Amantadine / Q000494:pharmacology; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D024882:Drug Resistance, Viral; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods; D006801:Humans; D009980:Influenza A virus / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests / Q000295:instrumentation / Q000379:methods; D015203:Reproducibility of Results; D014763:Viral Matrix Proteins / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects; D000086082:Viroporin Proteins
D000547:Amantadine; D000998:Antiviral Agents; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins