15N and 31P solid-state NMR study of transmembrane domain alignment of M2 protein of influenza A virus in hydrated cylindrical lipid bilayers confined to anodic aluminum oxide nanopores.
This communication reports the first example of a high resolution solid-state 15N 2D PISEMA NMR spectrum of a transmembrane peptide aligned using hydrated cylindrical lipid bilayers formed inside nanoporous anodic aluminum oxide (AAO) substrates. The transmembrane domain SSDPLVVA(A-15N)SIIGILHLILWILDRL of M2 protein from influenza A virus was reconstituted in hydrated 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine bilayers that were macroscopically aligned by a conventional micro slide glass support or by the AAO nanoporous substrate. 15N and 31P NMR spectra demonstrate that both the phospholipids and the protein transmembrane domain are uniformly aligned in the nanopores. Importantly, nanoporous AAO substrates may offer several advantages for membrane protein alignment in solid-state NMR studies compared to conventional methods. Specifically, higher thermal conductivity of aluminum oxide is expected to suppress thermal gradients associated with inhomogeneous radio frequency heating. Another important advantage of the nanoporous AAO substrate is its excellent accessibility to the bilayer surface for exposure to solute molecules. Such high accessibility achieved through the substrate nanochannel network could facilitate a wide range of structure-function studies of membrane proteins by solid-state NMR.
Chekmenev
Eduard Y
EY
The Center for Interdisciplinary Magnetic Resonance, National High Magnetic Field Laboratory (NHMFL), Tallahassee, FL 32310, USA. chekmene@magnet.fsu.edu
Hu
Jun
J
Gor'kov
Peter L
PL
Brey
William W
WW
Cross
Timothy A
TA
Ruuge
Andres
A
Smirnov
Alex I
AI
D000537:Aluminum Oxide; D009980:Influenza A virus / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010759:Phosphorus Isotopes; D011487:Protein Conformation; D014764:Viral Proteins / Q000737:chemistry*
D008051:Lipid Bilayers; D008565:Membrane Proteins; D009587:Nitrogen Isotopes; D010759:Phosphorus Isotopes; D014764:Viral Proteins; D000537:Aluminum Oxide