Lipid-protein correlations in nanoscale phospholipid bilayers determined by solid-state nuclear magnetic resonance

Aleksandra Kijac, Amy Y. Shih, Andrew J. Nieuwkoop, Klaus Schulten, Stephen G. Sligar, Chad M. Rienstra

Research output: Contribution to journalArticlepeer-review

Abstract

Nanodiscs are examples of discoidal nanoscale lipid-protein particles that have been extremely useful for the biochemical and biophysical characterization of membrane proteins. They are discoidal lipid bilayer fragments encircled and stabilized by two amphipathic helical proteins named membrane scaffolding protein (MSP), ∼10 nm in size. Nanodiscs are homogeneous, easily prepared with reproducible success, amenable to preparations with a variety of lipids, and stable over a range of temperatures. Here we present solid-state nuclear magnetic resonance (SSNMR) studies on lyophilized, rehydrated POPC Nanodiscs prepared with uniformly 13C-, 15N-labeled MSP1D1 (Δ1-11 truncated MSP). Under these conditions, by SSNMR we directly determine the gel-to-liquid crystal lipid phase transition to be at 3 ± 2 °C. Above this phase transition, the lipid 1H signals have slow transverse relaxation, enabling filtering experiments as previously demonstrated for lipid vesicles. We incorporate this approach into two- and three-dimensional heteronuclear SSNMR experiments to examine the MSP1D1 residues interfacing with the lipid bilayer. These 1H-13C and 1H-13C-13C correlation spectra are used to identify and quantify the number of lipid-correlated and solvent-exposed residues by amino acid type, which furthermore is compared with molecular dynamics studies of MSP1D1 in Nanodiscs. This study demonstrates the utility of SSNMR experiments with Nanodiscs for examining lipid-protein interfaces and has important applications for future structural studies of membrane proteins in physiologically relevant formulations.

Original languageEnglish (US)
Pages (from-to)9190-9198
Number of pages9
JournalBiochemistry
Volume49
Issue number43
DOIs
StatePublished - Nov 2 2010

ASJC Scopus subject areas

  • Biochemistry

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