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首页> 外文期刊>Biochemistry >Detection and quantification of asymmetric lipid vesicle fusion using deuterium NMR.
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Detection and quantification of asymmetric lipid vesicle fusion using deuterium NMR.

机译：使用氘核磁共振检测和定量不对称脂质囊泡融合。

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摘要

It is demonstrated that deuterium nuclear magnetic resonance (2H NMR) spectroscopy can be used to detect and to quantify fusion between anionic giant unilamellar vesicles (GUVs) and cationic small unilamellar vesicles (SUVs). The sensitivity to fusion relies on the conformational response of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) to changes in membrane surface electrostatic charge due to lipid mixing upon fusion. This conformational change is reported in the 2H NMR spectrum as a change in the quadrupolar splitting from choline-deuterated POPC. GUVs were composed of varying molar ratios of the anionic lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), plus cholesterol (CHOL), plus POPC. SUVs were composed of the cationic lipid 1,2-dioleoyloxy-3-(dimethylammonio)-propane (DODAP), plus POPC with or without 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE). Using a quantitative model that relates the 2H NMR quadrupolar splitting to the mole fractions of cationic, anionic, and neutral lipids in the vesicle membrane, it was possible to deduce the extent of fusion between the two oppositely-charged vesicle populations directly from the quadrupolar splitting. SUVs composed of DODAP + POPC + POPE (40/40/20) fused 100% with GUVs composed of POPC + CHOL + POPG (60/30/10). Removing POPE from the SUVs reduced the extent of fusion, as did reducing the POPG content of the GUVs.
机译：结果表明，氘核磁共振波谱（2H NMR）可用于检测和量化阴离子巨型单层囊泡（GUVs）和阳离子小型单层囊泡（SUVs）之间的融合。对融合的敏感性取决于1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸胆碱（POPC）对由于融合时脂质混合而引起的膜表面静电荷变化的构象响应。这种构象变化在2H NMR光谱中报告为胆碱氘代POPC在四极分裂中的变化。 GUV由阴离子脂质1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸甘油（POPG），胆固醇（CHOL）和POPC的不同摩尔比组成。 SUV由阳离子脂质1,2-二油酰氧基-3-（二甲基铵）-丙烷（DODAP）加上含或不含1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸乙醇胺（POPE）的POPC组成。使用将2H NMR四极分裂与囊膜中阳离子，阴离子和中性脂质的摩尔分数相关联的定量模型，可以直接从四极分裂推论两个带相反电荷的囊群之间的融合程度。由DODAP + POPC + POPE（40/40/20）组成的SUV与由POPC + CHOL + POPG（60/30/10）组成的GUV融合100％。从SUV上去除POPE减少了融合的程度，减少了GUV的POPG含量也是如此。

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