Membrane curvature change induced by an antimicrobial peptide detected by 31P exchange NMR.
The influence of an antimicrobial peptide, protegrin-1 (PG-1), on the curvature and lateral diffusion coefficient (D(L)) of phosphocholine bilayers is investigated using one- (1D) and two-dimensional (2D) (31)P exchange NMR. The experiments utilize the fact that lipid lateral diffusion over the curved surface of vesicles changes the molecular orientation and thus the (31)P chemical shift anisotropy. This reorientation is manifested in 2D spectra as off-diagonal intensities and in 1D stimulated-echo experiments as reduced echo heights. The 2D spectra give information on the reorientation-angle distribution while the decay of the stimulated-echo intensity, which closely tracks the second-order correlation function in our experiments, yields the correlation times of the reorientation. The relationships among the 2D exchange spectra, stimulated-echo intensities, the correlation function, and reorientation-angle distributions are analyzed in detail. In the absence of PG-1, both dilaurylphosphotidylcholine (DLPC) and palmitoyloleoylphosphatidylcholine (POPC) vesicles show biexponential decays of the stimulated-echo intensities to equilibrium values of 0.20-0.25, suggesting that the curvature of the lipid vesicles has a bimodal distribution. The addition of PG-1 to DLPC vesicles increased the decay time constants, indicating that D(L) decreases due to peptide binding. In contrast, the addition of PG-1 to POPC vesicles decreased the decay constants by three to fivefold, indicating that the POPC vesicles are fragmented into smaller vesicles. On the basis of the changes in D(L) and the decay constants, we estimate that the radius of the POPC vesicles decreases by threefold due to PG-1 binding. Simulations of the 2D exchange spectra yielded quantitative reorientation-angle distributions that are consistent with the bimodal distributions of the vesicle curvature and the effects of the peptide on the two types of lipid bilayers. Thus, (31)P exchange NMR provides useful insights into the membrane morphological changes induced by this antimicrobial peptide.
Marasinghe
P A B
PA
Department of Chemistry, Iowa State University, Ames, Iowa 50011, USA.
Buffy
J J
JJ
Schmidt-Rohr
K
K
Hong
M
M
D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008567:Membranes, Artificial; D010713:Phosphatidylcholines / Q000737:chemistry*; D010759:Phosphorus Isotopes; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013997:Time Factors
D023181:Antimicrobial Cationic Peptides; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D010759:Phosphorus Isotopes; D011506:Proteins; C094880:protegrin-1; C015153:1,2-dilauroylphosphatidylcholine; C028694:1-palmitoyl-2-oleoylphosphatidylcholine