tetano
Editor, Senior Moderator
J Virol. 2013 Jul 24. [Epub ahead of print]
Detection of proton movement directly across viral membranes to identify novel influenza M2 inhibitors.
Sulli C, Banik SS, Schilling J, Moser A, Xiang X, Payne R, Wanless A, Willis SH, Paes C, Rucker JB, Doranz BJ.
Source
Integral Molecular, Inc. 3711 Market St, Suite 900, Philadelphia, PA 19104.
Abstract
The influenza M2 protein is a well-validated yet underexploited proton-selective ion channel essential for influenza virus infectivity. Because M2 is a toxic viral ion channel, existing M2 inhibitors have been discovered through live virus inhibition or medicinal chemistry rather than M2-targeted high-throughput screening (HTS), and direct measurement of its activity has been limited to live cells or reconstituted lipid bilayers. Here we describe a cell-free ion channel assay in which M2 ion channels are incorporated into virus-like particles (VLPs) and proton conductance is measured directly across the viral lipid bilayer, detecting changes in membrane potential, ion permeability, and ion channel function. Using this approach, we identified 19 M2-specific inhibitors, including two novel chemical scaffolds that inhibit both M2 function and influenza virus infectivity. Counter-screening for non-specific disruption of viral bilayer ion permeability also identified a broad-spectrum anti-viral compound that acts by disrupting the integrity of the viral membrane. In addition to its application to M2 and potentially other ion channels, this technology enables direct measurement of the electrochemical and biophysical characteristics of viral membranes.
PMID:
23885079
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23885079
Detection of proton movement directly across viral membranes to identify novel influenza M2 inhibitors.
Sulli C, Banik SS, Schilling J, Moser A, Xiang X, Payne R, Wanless A, Willis SH, Paes C, Rucker JB, Doranz BJ.
Source
Integral Molecular, Inc. 3711 Market St, Suite 900, Philadelphia, PA 19104.
Abstract
The influenza M2 protein is a well-validated yet underexploited proton-selective ion channel essential for influenza virus infectivity. Because M2 is a toxic viral ion channel, existing M2 inhibitors have been discovered through live virus inhibition or medicinal chemistry rather than M2-targeted high-throughput screening (HTS), and direct measurement of its activity has been limited to live cells or reconstituted lipid bilayers. Here we describe a cell-free ion channel assay in which M2 ion channels are incorporated into virus-like particles (VLPs) and proton conductance is measured directly across the viral lipid bilayer, detecting changes in membrane potential, ion permeability, and ion channel function. Using this approach, we identified 19 M2-specific inhibitors, including two novel chemical scaffolds that inhibit both M2 function and influenza virus infectivity. Counter-screening for non-specific disruption of viral bilayer ion permeability also identified a broad-spectrum anti-viral compound that acts by disrupting the integrity of the viral membrane. In addition to its application to M2 and potentially other ion channels, this technology enables direct measurement of the electrochemical and biophysical characteristics of viral membranes.
PMID:
23885079
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/23885079