Preparation of artificial vesicles having an L-alanine uptake activity which requires NADH as energy source

Abstract

The vesicles (proteoliposomes) were artificially prepared by sonic oscillation from hydrophobic protein, which was isolated from cytoplasmic membrane fraction of Bacillus subtilis W-23, and phospholipids or lipids, which were isolated from different organims; phospholipids from hen-egg yolk and soybean, and lipids from B . subtilis W-23. In the vesicles, remarkable activity of L-alanine uptake was demonstrated in the presence of NADH or ascorbate-phenazine methosulfate as an energy source. The uptake activity was temperature-dependent and inhibited by respiration inhibitor (KCN) and uncoupler (ClCCP). Replacement experiment was also performed.