Siegel D P, Ware B R
Biophys J. 1980 Apr;30(1):159-72. doi: 10.1016/S0006-3495(80)85084-3.
Using the technique of electrophoretic light scattering, we have measured the electrophoretic mobilities of synaptic vesicles and synaptosomal plasma membranes isolated from guinea-pig cerebral cortex. The electrophoretic mobility of synaptic vesicles is slightly greater than that of synaptosomal plasma membranes. Ca+2 and Mg+2 reduced the mobility of both species to the same extent at physiologically relevant concentrations (0-1 mM) and near-physiologic ionic strength. The extent of the reduction was not large (approximately 6% for synaptic vesicles in the presence of 100 mM KCl) at 1 mM divalent cation concentrations. At concentrations of approximately 2 mM and higher, Ca+2 reduced the mobility of synaptic vesicles more than did Mg/2. A similar but much smaller effect was observed in the case of synaptosomal plasma membranes. The addition of 1 mM Mg+2-ATP had no effect upon synaptic vesicle mobility either in the presence or absence of the ionophores nigericin or valinomycin. These data, together with earlier work (Siegel et al., 1978, Biophys. J. 22:341-346), demonstrate that substantial reduction of the average electrostatic surface charge density is not the most important role of divalent cations in promoting close approach of secretory granules and secretory cell membranes, and that it is certainly not the Ca+2-specific step in exocytosis.
利用电泳光散射技术,我们测量了从豚鼠大脑皮层分离出的突触小泡和突触体细胞膜的电泳迁移率。突触小泡的电泳迁移率略高于突触体细胞膜。在生理相关浓度(0 - 1 mM)和接近生理离子强度下,Ca²⁺和Mg²⁺以相同程度降低了两者的迁移率。在1 mM二价阳离子浓度下,迁移率降低的幅度不大(在100 mM KCl存在下,突触小泡约为6%)。在约2 mM及更高浓度时,Ca²⁺比Mg²⁺更能降低突触小泡的迁移率。在突触体细胞膜的情况下也观察到了类似但小得多的效应。添加1 mM Mg²⁺ - ATP,无论是否存在尼日利亚菌素或缬氨霉素离子载体,对突触小泡迁移率均无影响。这些数据,连同早期的研究工作(Siegel等人,1978年,《生物物理杂志》22:341 - 346)表明,二价阳离子在促进分泌颗粒与分泌细胞膜紧密靠近方面,大幅降低平均静电表面电荷密度并非最重要的作用,而且这肯定不是胞吐作用中Ca²⁺特异性的步骤。