Issue 2, 2009

Cholesterol containing model membranes studied by multinuclear solid state NMR spectroscopy

Abstract

Ternary phase diagrams have been shown to be very useful in understanding the phenomena of lipid rafts. Recently a comprehensive ternary phase diagram for DPPC-d62/DOPC/Chol using static 2H-NMR has been reported [S. L. Veatch, O. Soubias, S. L. Keller, and K. Gawrisch Proceedings of the National Academy of Sciences of the United States of America, 2007, 104, 17650–17655]. However, it is often difficult to interpret these ternary phase diagrams as there are many possible ambiguities, in particular, the extent of the gel(so)–Lo–Lα(ld) three phase region and the clear demarcation between the gel(so) and Lo phase. Here we examine the canonical model lipid raft system by a multinuclear approach using 2H static and 13C, 31P Magic Angle Spinning (MAS) NMR. By using deuterated DPPC (DPPC-d62) it is possible to examine changes in the methylene chain order parameters of the saturated lipid whilst changing the ratio of saturated to unsaturated phospholipid. From these data it is possible to propose the existence at low temperatures (<20 °C) of a disrupted or disordered gel-type phase at ratios as high as 1:1:1 DOPC/DPPC/Chol, in equilibrium with fluid phases. The presence of a gel-type phase was also confirmed by 13C and 31P MAS NMR. This disrupted gel-type phase has different properties in terms of axial rotation than that of the liquid ordered phase (Lo). We have also shown that it is possible to observe different quadrupolar splittings for the inequivalent sn-1 and sn-2 methyl groups in a gel phase. We have concluded that this is not always indicative of a liquid-ordered lamellar Lo phase, as previously assumed. Above the melting point of the gel-type phase (∼20 °C), we observe a single fluid environment for both DPPC-d62 and Chol-d1 on the basis of the 2H-NMR spectra, which are an average of a fluid disordered (Lα) and an Lo phase. In this fluid regime, the order parameters for the 1:1:1 system DPPC-d62/DOPC/Chol are close to that of the binary (1:1) DPPC-d62:Chol system, indicating non-random distribution of the DPPC-d62. Increasing the ratio of DOPC to DPPC, from 1:1:1 to 1.5:1:1 to 2:1:1 in the ternary system, causes an increase in chain length of the DPPC-d62 in the fluid phase.

Graphical abstract: Cholesterol containing model membranes studied by multinuclear solid state NMR spectroscopy

Supplementary files

Article information

Article type
Paper
Submitted
29 May 2008
Accepted
02 Oct 2008
First published
06 Nov 2008

Soft Matter, 2009,5, 369-378

Cholesterol containing model membranes studied by multinuclear solid state NMR spectroscopy

J. A. Clarke, J. M. Seddon and R. V. Law, Soft Matter, 2009, 5, 369 DOI: 10.1039/B809139K

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