Mühleisen M, Probst W, Hayashi K, Rahmann H
Jpn J Exp Med. 1983 Apr;53(2):103-7.
Ca2+-binding to liposomes was investigated by means of equilibrium dialysis using 45Ca as tracer. Liposomes composed of various lipids (phospholipids: phosphatidylcholine, -serine, -inositol, -inositol-3, 4-bisphosphate; glycosphingolipids: ganglioside mixture from bovine brain; isolated ganglioside species GM1, GD1a, GT1b; cholesterol) in ratios resembling those of natural cell surface membranes were prepared by the detergent dialysis method. Ca2+ was applied in concentrations ranging from 5 microM to 0.15 mM. Pure phosphatidylcholine liposomes showed only poor Ca2+-binding affinities; however, negatively charged liposomes bound Ca2+ in a rather distinct and specific manner. Ca2+-binding to liposomes containing phosphatidylinositol-3,4-bisphosphate in their bilayers was highest. All the liposomes (except those containing phosphatidylinositol-3,4-bisphosphate) showed no simple Ca2+-binding characteristics, that is, continuous Ca2+-binding was interrupted at a concentration of about 50 microM of total Ca2+ in each Ca2+-binding plot, implying that a certain amount (10%-45%) of previously bound Ca2+ was released. These results are discussed with regard to the different possibilities on the molecular basis of physico-chemical processes as well as in relation to their possible functional role at the process of neuronal synaptic transmission.
以(^{45}Ca)作为示踪剂,通过平衡透析法研究了钙离子与脂质体的结合情况。采用去污剂透析法制备了由各种脂质(磷脂:磷脂酰胆碱、磷脂酰丝氨酸、磷脂酰肌醇、磷脂酰肌醇 - 3,4 - 二磷酸;糖鞘脂:牛脑来源的神经节苷脂混合物;分离得到的神经节苷脂种类GM1、GD1a、GT1b;胆固醇)按类似于天然细胞表面膜的比例组成的脂质体。钙离子的应用浓度范围为(5)微摩尔/升至(0.15)毫摩尔/升。纯磷脂酰胆碱脂质体显示出较差的钙离子结合亲和力;然而,带负电荷的脂质体以相当独特和特异的方式结合钙离子。双层中含有磷脂酰肌醇 - 3,4 - 二磷酸的脂质体对钙离子的结合能力最强。所有脂质体(除了含有磷脂酰肌醇 - 3,4 - 二磷酸的脂质体)均未表现出简单的钙离子结合特性,即在每个钙离子结合曲线中,当总钙离子浓度约为(50)微摩尔/升时,连续的钙离子结合会中断,这意味着先前结合的一定量((10%-45%))的钙离子会被释放。从物理化学过程的分子基础的不同可能性以及它们在神经元突触传递过程中可能的功能作用方面对这些结果进行了讨论。