Dilute spin-exchange assignment of solid-state NMR spectra of oriented proteins: acetylcholine M2 in bilayers.
The assignment of amide resonances in the two-dimensional PISEMA (Polarization Inversion with Spin Exchange at the Magic Angle) spectrum of uniformly 15N labeled M2 peptide corresponding to the channel-lining segment of the acetylcholine receptor in oriented phospholipid bilayers is described. The majority of the resonances were assigned through comparisons with spectra from selectively 15N labeled recombinant peptides and specifically 15N labeled synthetic peptides. Some resonances were assigned to specific amino acid residues by means of homonuclear 15N spin-exchange spectroscopy. A modification to the conventional spin-exchange pulse sequence that significantly shortens the length of the experiments by combining the intervals for 15N spin-exchange and 1H magnetization recovery is described.
Marassi
F M
FM
Department of Chemistry, University of Pennsylvania, Philadelphia 19104, USA.
Gesell
J J
JJ
Valente
A P
AP
Kim
Y
Y
Oblatt-Montal
M
M
Montal
M
M
Opella
S J
SJ
D000595:Amino Acid Sequence; D008051:Lipid Bilayers; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000295:instrumentation / Q000379:methods; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation; D043585:Receptor, Muscarinic M2; D011976:Receptors, Muscarinic / Q000737:chemistry*
D008051:Lipid Bilayers; D009587:Nitrogen Isotopes; D010446:Peptide Fragments; D043585:Receptor, Muscarinic M2; D011976:Receptors, Muscarinic