Schmidt C F, Lichtenberg D, Thompson T E
Biochemistry. 1981 Aug 4;20(16):4792-7. doi: 10.1021/bi00519a041.
The time course of the size transformation of sonicated small unilamellar dipalmitoylphosphatidylcholine vesicles at 23 degrees C has been followed with (31)P and (1)H nuclear magnetic resonance (NMR) spectroscopy. Comparison of these results with turbidity measurements indicates that vesicle aggregation, monitored by turbidity, and size transformation, followed by NMR, occur on distinctly different time scales. For lipid concentrations in the 5-50 mM range, aggregation takes place on a time scale of minutes, whereas size transformation takes many hours. Aggregation, unlike size transformation, can be reversed by increasing the temperature above the phospholipid phase transition temperature. Analysis of the (31)P NMR line shapes provides evidence for a model in which the small vesicles transform into the product vesicles [characterized by Schullery, S.E., Schmidt, C.F., Felgner, P., Tillack, T.W., & Thompson, T.E. (1980) Biochemistry 19, 3919] without producing significant amounts of intermediate-size vesicles. Kinetic analysis indicates that the size transformation is apparently second order. (1)H NMR data indicate that the rate of transformation is decreased if trivalent ions are added to the dispersions and also if the temperature is periodically increased above the transition temperature. Analysis of the latter experiment provides some evidence that vesicle aggregation is a necessary precursor to size transformation. It was also found that increasing the average vesicle size decreases the extent of transformation.
利用³¹P和¹H核磁共振(NMR)光谱跟踪了在23℃下超声处理的小单层二棕榈酰磷脂酰胆碱囊泡尺寸转变的时间进程。将这些结果与浊度测量结果进行比较表明,通过浊度监测的囊泡聚集和通过NMR跟踪的尺寸转变发生在明显不同的时间尺度上。对于5 - 50 mM范围内的脂质浓度,聚集发生在几分钟的时间尺度上,而尺寸转变则需要数小时。与尺寸转变不同,聚集可以通过将温度升高到磷脂相变温度以上而逆转。对³¹P NMR线形的分析为一个模型提供了证据,在该模型中,小囊泡转变成产物囊泡[由Schullery, S.E., Schmidt, C.F., Felgner, P., Tillack, T.W., & Thompson, T.E.(1980年)在《生物化学》19卷,3919页中描述],而不会产生大量中等尺寸的囊泡。动力学分析表明,尺寸转变显然是二级反应。¹H NMR数据表明,如果向分散体中添加三价离子,以及如果温度周期性地升高到转变温度以上,转变速率会降低。对后一个实验的分析提供了一些证据,表明囊泡聚集是尺寸转变的必要前提。还发现增加平均囊泡尺寸会降低转变程度。