Artificial cell based on lipid hollow polyelectrolyte microcapsules: channel reconstruction and membrane potential measurement.

摘要

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.