Transmembrane domain of M2 protein from influenza A virus studied by solid-state (15)N polarization inversion spin exchange at magic angle NMR.
The M2 protein from the influenza A virus forms a proton channel in the virion that is essential for infection. This tetrameric protein appears to form a four-helix bundle spanning the viral membrane. Here the solid-state NMR method, 2D polarization inversion spin exchange at magic angle (PISEMA), has been used to obtain multiple constraints from specifically amino acid-labeled samples. The improvement of spectral resolution from 2D PISEMA over 1D methods and 2D separated local field methods is substantial. The reliability of the method is validated by comparison of anisotropic chemical shift and heteronuclear dipolar interactions from single site labeled samples. The quantitative interpretation of the high-resolution constraints confirms the helix tilt to be within the range of previous experimental determinations (32 degrees -38 degrees ). The binding of the channel inhibitor, amantadine, results in no change in the backbone structure at position Val(27,28), which is thought to be a potential binding site for the inhibitor.
Song
Z
Z
National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida 32306-4005, USA.
Kovacs
F A
FA
Wang
J
J
Denny
J K
JK
Shekar
S C
SC
Quine
J R
JR
Cross
T A
TA
D000595:Amino Acid Sequence; D016880:Anisotropy; D007473:Ion Channels / Q000737:chemistry; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D010446:Peptide Fragments / Q000737:chemistry*; D011487:Protein Conformation; D015203:Reproducibility of Results; D014763:Viral Matrix Proteins / Q000737:chemistry*; D000086082:Viroporin Proteins
D007473:Ion Channels; D008051:Lipid Bilayers; D009587:Nitrogen Isotopes; D010446:Peptide Fragments; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins