Tiourina O P, Radtchenko I, Sukhorukov G B, Möhwald H
Max Planck Institute of Colloids and Interfaces, Golm/Potsdam, D-14476, Germany.
J Membr Biol. 2002 Nov 1;190(1):9-16. doi: 10.1007/s00232-002-1018-4.
A new model system as an artificial cell based on hollow polyelectrolyte microcapsules (HPM), fabricated by step-wise adsorption of polyelectrolytes and phospholipids, was proposed. To demonstrate the feasibility of the functioning of HPM coated with lipid layers as a model biosystem, the membrane permeability for ions and the channel reconstruction were examined by measuring the membrane potential as one of the most important cell parameters. The membrane potential was measured by confocal microscopy using the potentiometric fluorescent dye tetramethylrhodamine ethyl ester perchlorate (TMRE). The influence of the lipid composition (phosphatidylcholine--DPPC-HPM, phosphatidylcholine/phosphatidic acid 9/1-(DPPC/DPPA)-HPM) and metal cations (Na+, K+, Ca2+) on the membrane potential was shown. Addition of DPPA was found to lead to an increase of the negative membrane potential value. To investigate the ion-transport activity of the lipid-HPM, the ion channel-forming peptides valinomycin and gramicidin were used. The selectivity of the valinomycin and gramicidin channels for K+ and for K+, and Na+ ions, respectively, and the tolerance for Ca2+ ions is evidence that lipid-HPM functions as an artificial cell.
提出了一种基于中空聚电解质微胶囊(HPM)的新型人工细胞模型系统,该系统通过逐步吸附聚电解质和磷脂制备而成。为了证明涂有脂质层的HPM作为模型生物系统发挥功能的可行性,通过测量作为最重要细胞参数之一的膜电位,研究了离子的膜通透性和通道重建。使用电位荧光染料四甲基罗丹明乙酯高氯酸盐(TMRE)通过共聚焦显微镜测量膜电位。研究了脂质组成(磷脂酰胆碱-DPPC-HPM、磷脂酰胆碱/磷脂酸9/1-(DPPC/DPPA)-HPM)和金属阳离子(Na+、K+、Ca2+)对膜电位的影响。发现添加DPPA会导致膜电位负值增加。为了研究脂质-HPM的离子转运活性,使用了离子通道形成肽缬氨霉素和短杆菌肽。缬氨霉素和短杆菌肽通道分别对K+以及对K+和Na+离子的选择性,以及对Ca2+离子的耐受性,证明脂质-HPM起到了人工细胞的作用。