Berrier C, Besnard M, Ajouz B, Coulombe A, Ghazi A
Laboratoire des Biomembranes, URA CNRS 1116, Bât. 432, Université Paris-Sud, 91405 Orsay, France.
J Membr Biol. 1996 May;151(2):175-87. doi: 10.1007/s002329900068.
Mechanosensitive ion channels from Escherichia coli were studied in giant proteoliposomes reconstituted from an inner membrane fraction, or in giant round cells in which the outer membrane and the cell wall had been disrupted by a lysozyme-EDTA treatment and a mild osmotic shock. Patch-clamp experiments revealed the presence in these two preparations of an array of different conductances (100 to 2,300 pS in 0.1 M KCl) activated by stretch. The electrical activity induced by stretch in the native membrane was complex, due to the activation of several different conductances. In contrast, patches of proteoliposomes generally contained clusters of identical conductances, which differed from patch to patch. These experiments are consistent with the notion that these different conductances correspond to different proteins in the plasma membrane of E. coli, which segregate into clusters of identical channels on dilution involved in reconstitution in proteoliposomes. These conductances could be grouped into three subfamilies of poorly selective channels. In both preparations, the higher the conductance, the higher was the negative pressure needed for activation. We discuss the putative role of these channels as parts of a multicomponent osmoregulatory system.
利用从内膜组分重构的巨大蛋白脂质体,或通过溶菌酶 - 乙二胺四乙酸处理和轻度渗透压休克破坏外膜和细胞壁的巨大圆形细胞,对来自大肠杆菌的机械敏感离子通道进行了研究。膜片钳实验表明,在这两种制剂中存在一系列由拉伸激活的不同电导(在0.1 M KCl中为100至2300 pS)。由于几种不同电导的激活,天然膜中由拉伸诱导的电活动很复杂。相比之下,蛋白脂质体膜片通常包含相同电导的簇,不同膜片之间存在差异。这些实验与以下观点一致,即这些不同的电导对应于大肠杆菌质膜中的不同蛋白质,在重构到蛋白脂质体中稀释时,它们会分离成相同通道的簇。这些电导可分为选择性较差的通道的三个亚家族。在这两种制剂中,电导越高,激活所需的负压就越高。我们讨论了这些通道作为多组分渗透调节系统一部分的假定作用。