Journal
BIOCHEMISTRY-MOSCOW
Volume 74, Issue 7, Pages 756-765Publisher
MAIK NAUKA/INTERPERIODICA/SPRINGER
DOI: 10.1134/S0006297909070086
Keywords
nanodisc; apolipoprotein; high-density lipoprotein particle; membrane protein; membrane-active peptide; model membranes; NMR spectroscopy
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Funding
- Russian Federation President's [MK-6346.2008.4, SS-1061.2008.4]
- Russian Academy of Sciences
- Russian Foundation for Basic Research [06-04-49413, 08-04-01442]
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High-resolution NMR is shown to be applicable for investigation of membrane proteins and membrane-active peptides embedded into lipid-protein nanodiscs (LPNs). N-15-Labeled K+-channel from Streptomyces lividans (KcsA) and the antibiotic antiamoebin I from Emericellopsis minima (Aam-I) were embedded in LPNs of different lipid composition. Formation of stable complexes undergoing isotropic motion in solution was confirmed by size-exclusion chromatography and P-31-NMR spectroscopy. The 2D H-1-N-15-correlation spectra were recorded for KcsA in the complex with LPN containing DMPC and for Aam-I in LPNs based on DOPG, DLPC, DMPC, and POPC. The spectra recorded were compared with those in detergent-containing micelles and small bicelles commonly used in high-resolution NMR spectroscopy of membrane proteins. The spectra recorded in LPN environments demonstrated similar signal dispersion but significantly increased H-1(N) line width. The spectra of Aam-I embedded in LPNs containing phosphatidylcholine showed significant selective line broadening, thus suggesting exchange process(es) between several membrane-bound states of the peptide. N-15 relaxation rates were measured to obtain the effective rotational correlation time of the Aam-I molecule. The obtained value (similar to 40 nsec at 45A degrees C) is indicative of additional peptide motions within the Aam-I/LPN complex.
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