A high-resolution solid-state NMR approach for the structural studies of bicelles - PubMed (original) (raw)

A high-resolution solid-state NMR approach for the structural studies of bicelles

Sergey Dvinskikh et al. J Am Chem Soc. 2006.

Abstract

Bicelles are increasingly being used as membrane mimicking systems in NMR experiments to investigate the structure of membrane proteins. In this study, we demonstrate the effectiveness of a 2D solid-state NMR approach that can be used to measure the structural constraints, such as heteronuclear dipolar couplings between 1H, 13C, and 31P nuclei, in bicelles without the need for isotopic enrichment. This method does not require a high radio frequency power unlike the presently used rotating-frame separated-local-field (SLF) techniques, such as PISEMA. In addition, multiple dipolar couplings can be measured accurately, and the presence of a strong dipolar coupling does not suppress the weak couplings. High-resolution spectra obtained from magnetically aligned DMPC:DHPC bicelles even in the presence of peptides suggest that this approach will be useful in understanding lipid-protein interactions that play a vital role in shaping up the function of membrane proteins.

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Figures

Figure 1

Figure 1

13C chemical shift spectrum (left), 2D spectrum (middle) that correlates the 13C chemical shift (vertical dimension) and 1H-13C dipolar coupling (horizontal dimension) and 1H-13C dipolar coupling slices (right) of 3.5:1 DMPC:DHPC bicelles obtained from a Varian/Chemagnetics 400 MHz solid-state NMR spectrometer. The pulse sequence given in the Supporting Information was used. 64 scans were accumulated for each of 200 points in _t_1 dimensions with the increment time of 384 _μ_s and recycle delay of 5s. 1H rf field strength of 31 kHz were used during _t_1 evolution. Contact time for CP transfer was set to 3.0 ms.

Figure 2

Figure 2

(A) 1H-13C dipolar coupling extracted from 2D spectra from 3.5:1 DMPC:DHPC bicelles with 0 (bottom), 0.5 (middle), and 2 (top) mole % MSI-78. (B) The order parameter profile for DMPC determined from the dipolar coupling values.

Figure 3

Figure 3

(A) A 2D correlation of 13C chemical shift and and 13C-31P dipolar coupling (y-axis) with the 1H chemical shift and 1H-31P dipolar coupling (x-axis) of 3.5:1 DMPC:DHPC bicelles (left). The 1H-31P dipolar split peaks of α, β, g2 and g3 are connected by a green line. (B) The measured 1H-31P (blue) and 13C-31P (red) dipolar couplings in DMPC lipid molecule are highlighted.

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