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Lipid-protein nanodiscs: Possible application in high-resolution NMR investigations of membrane proteins and membrane-active peptides

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Abstract

High-resolution NMR is shown to be applicable for investigation of membrane proteins and membrane-active peptides embedded into lipid-protein nanodiscs (LPNs). 15N-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 31P-NMR spectroscopy. The 2D 1H-15N-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 1HN 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. 15N relaxation rates were measured to obtain the effective rotational correlation time of the Aam-I molecule. The obtained value (∼40 nsec at 45°C) is indicative of additional peptide motions within the Aam-I/LPN complex.

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Abbreviations

Aam-I:

antibiotic antiamoebin I from Emericellopsis minima

apoA-1:

apolipoprotein A-1

CRINEPT:

cross-correlated relaxation-enhanced polarization

1D:

one-dimensional

2D:

two-dimensional

DDM:

β-dodecyl maltoside

DHPC:

dihexanoyl phosphatidylcholine

DLPC:

dilauroyl phosphatidylcholine

DLPG:

dilauroyl phosphatidylglycerol

DMPC:

dimyristoyl phosphatidylcholine

DOPC:

dioleoyl phosphatidylcholine

DOPE:

dioleoyl phosphatidylethanolamine

DOPG:

dioleoyl phosphatidylglycerol

HMQC:

heteronuclear multiple quantum correlation

HSQC:

heteronuclear single quantum correlation

INEPT:

insensitive nuclei enhanced by polarization transfer

KcsA:

K+-channel from Streptomyces lividans

LMPC:

lysomyristoyl phosphatidylcholine

LMPG:

lysomyristoyl phosphatidylglycerol

LPN:

lipid-protein nanodisc

MP:

membrane protein

MSP:

fragment 44-243 of human apolipoprotein A1 (membrane scaffold protein)

POPC:

palmitoyloleoyl phosphatidylcholine

R St :

hydrodynamic radius of a particle, Stokes radius

TM:

transmembrane

TROSY:

transverse relaxation-optimized spectroscopy

Δν:

half-peak width of NMR band

ηXY :

rate of cross-correlation between dipole-dipole and chemical shift anisotropy relaxation of the 15N nucleus

τR :

effective rotational correlation time

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117997, Moscow, Russia

    Z. O. Shenkarev, E. N. Lyukmanova, O. I. Solozhenkin, I. E. Gagnidze, O. V. Nekrasova, V. V. Chupin, A. A. Tagaev, Z. A. Yakimenko, T. V. Ovchinnikova, M. P. Kirpichnikov & A. S. Arseniev

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  1. Z. O. Shenkarev
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  2. E. N. Lyukmanova
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  3. O. I. Solozhenkin
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  4. I. E. Gagnidze
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  5. O. V. Nekrasova
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  6. V. V. Chupin
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  7. A. A. Tagaev
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  8. Z. A. Yakimenko
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  9. T. V. Ovchinnikova
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  11. A. S. Arseniev
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Correspondence to Z. O. Shenkarev.

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Original Russian Text © Z. O. Shenkarev, E. N. Lyukmanova, O. I. Solozhenkin, I. E. Gagnidze, O. V. Nekrasova, V. V. Chupin, A. A. Tagaev, Z. A. Yakimenko, T. V. Ovchinnikova, M. P. Kirpichnikov, A. S. Arseniev, 2009, published in Biokhimiya, 2009, Vol. 74, No. 7, pp. 933–945.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM09-026, May 10, 2009.

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Shenkarev, Z.O., Lyukmanova, E.N., Solozhenkin, O.I. et al. Lipid-protein nanodiscs: Possible application in high-resolution NMR investigations of membrane proteins and membrane-active peptides. Biochemistry Moscow 74, 756–765 (2009). https://doi.org/10.1134/S0006297909070086

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  • DOI: https://doi.org/10.1134/S0006297909070086

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