Nitrogen-14 solid-state NMR spectroscopy of aligned phospholipid bilayers to probe peptide-lipid interaction and oligomerization of membrane associated peptides.
Characterization of the oligomerization of membrane-associated peptides is important to understand the folding and function of biomolecules like antimicrobial peptides, fusion peptides, amyloid peptides, toxins, and ion channels. However, this has been considered to be very difficult, because the amphipathic properties of the constituents of the cell membrane pose tremendous challenges to most commonly used biophysical techniques. In this study, we present the application of a simple (14)N solid-state NMR spectroscopy of aligned model membranes containing a phosphatidyl choline lipid to investigate the oligomerization of membrane-associated peptides. Since the near-symmetric nature of the choline headgroup of a phosphocholine lipid considerably reduces the (14)N quadrupole coupling, there are significant practical advantages in using (14)N solid-state NMR experiments to probe the interaction of peptide or protein with the surface of model membranes. Experimental results for several membrane-associated peptides are presented in this paper. Our results suggest that the experimentally measured (14)N quadrupole splitting of the lipid depends on the peptide-induced changes in the electrostatic potential of the lipid bilayer surface and therefore on the nature of the peptide, peptide-membrane interaction, and peptide-peptide interaction. It is inferred that the membrane orientation and oligomerization of the membrane-associated peptides can be measured using (14)N solid-state NMR spectroscopy.
Ramamoorthy
Ayyalusamy
A
Biophysics and Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109-1055, USA. ramamoor@umich.edu
Lee
Dong-Kuk
DK
Santos
Jose S
JS
Henzler-Wildman
Katherine A
KA
D008051:Lipid Bilayers / Q000737:chemistry*; D008055:Lipids / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods* / Q000592:standards; D008567:Membranes, Artificial; D015394:Molecular Structure; D009584:Nitrogen / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry*; D010758:Phosphorus / Q000737:chemistry; D012015:Reference Standards
D008051:Lipid Bilayers; D008055:Lipids; D008567:Membranes, Artificial; D010455:Peptides; D010743:Phospholipids; D010758:Phosphorus; D009584:Nitrogen