Sun S T, Hsang C C, Day E P, Ho J T
Biochim Biophys Acta. 1979 Oct 19;557(1):45-52. doi: 10.1016/0005-2736(79)90088-9.
Dynamic light scattering has been used to study the temperature dependence of Ca2+-induced fusion of phosphatidylserine vesicles and mixed vesicles containing phosphatidylserine and different phosphatidylcholines. The final vesicle size after Ca2+ and EDTA incubation serves as a measure of the extent of fusion. With phosphatidylserine vesicles, the extent of fusion shows a sharp maximum at an incubation temperature which depends on the Ca2+ concentration between 0.8 and 2 mM. The shift in the fusion peak temperature with Ca2+ concentration is similar to the typical shift in the phase transition temperature with divalent cation concentration in acidic phospholipids. The results suggest a direct correlation between the fusion peak temperature and the phase transition temperature in the presence of Ca2+ prior to fusion. With mixed vesicles containing up to 33% of a phosphatidylcholine in at least 2 mM Ca2+, the extent of fusion as a function of incubation temperature also shows a maximum. The fusion peak temperature is essentially independent of the quantity and type of phosphatidylcholine and the Ca2+ concentration, and identical to that with pure phosphatidylserine in excess Ca2+. The results imply that Ca2+- induced molecular segregation occurs first, and fusion subsequently takes place between pure phosphatidylserine domains.
动态光散射已被用于研究钙离子诱导的磷脂酰丝氨酸囊泡以及含有磷脂酰丝氨酸和不同磷脂酰胆碱的混合囊泡融合的温度依赖性。钙离子和乙二胺四乙酸孵育后的最终囊泡大小可作为融合程度的一个衡量指标。对于磷脂酰丝氨酸囊泡,融合程度在一个孵育温度下呈现出尖锐的最大值,该温度取决于0.8至2 mM之间的钙离子浓度。融合峰温度随钙离子浓度的变化类似于酸性磷脂中相变温度随二价阳离子浓度的典型变化。结果表明,在融合前钙离子存在的情况下,融合峰温度与相变温度之间存在直接关联。对于含有高达33%磷脂酰胆碱且钙离子浓度至少为2 mM的混合囊泡,融合程度作为孵育温度的函数也呈现出一个最大值。融合峰温度基本上与磷脂酰胆碱的量和类型以及钙离子浓度无关,并且与过量钙离子存在下的纯磷脂酰丝氨酸的融合峰温度相同。结果表明,首先发生钙离子诱导的分子分离,随后在纯磷脂酰丝氨酸结构域之间发生融合。