Freezing Point Depression of Water in Phospholipid Membranes: A Solid-State NMR Study

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• 作者：Dong-Kuk Lee ; Byung Soo Kwon ; Ayyalusamy Ramamoorthy
• 刊名：Langmuir
• 出版年：2008
• 出版时间：December 2, 2008
• 年：2008
• 卷：24
• 期：23
• 页码：13598-13604
• 全文大小：176K
• 年卷期：v.24,no.23(December 2, 2008)
• ISSN：1520-5827

文摘

Lipid−water interaction plays an important role in the properties of lipid bilayers, cryoprotectants, and membrane-associated peptides and proteins. The temperature at which water bound to lipid bilayers freezes is lower than that of free water. Here, we report a solid-state NMR investigation on the freezing point depression of water in phospholipid bilayers in the presence and absence of cholesterol. Deuterium NMR spectra at different temperatures ranging from −75 to +10 °C were obtained from fully ²H₂O-hydrated POPC (1-palmitoyl-2-oleoylphosphatidylcholine) multilamellar vesicles (MLVs), prepared with and without cholesterol, to determine the freezing temperature of water and the effect of cholesterol on the freezing temperature of water in POPC bilayers. Our ²H NMR experiments reveal the motional behavior of unfrozen water molecules in POPC bilayers even at temperatures significantly below 0 °C and show that the presence of cholesterol further lowered the freezing temperature of water in POPC bilayers. These results suggest that in the presence of cholesterol the fluidity and dynamics of lipid bilayers can be retained even at very low temperatures as exist in the liquid crystalline phase of the lipid. Therefore, bilayer samples prepared with a cryoprotectant like cholesterol should enable the performance of multidimensional solid-state NMR experiments to investigate the structure, dynamics, and topology of membrane proteins at a very low temperature with enhanced sample stability and possibly a better sensitivity. Phosphorus-31 NMR data suggest that lipid bilayers can be aligned at low temperatures, while ¹⁵N NMR experiments demonstrate that such aligned samples can be used to enhance the signal-to-noise ratio of ¹⁵N chemical shift spectra of a 37-residue human antimicrobial peptide, LL-37.