用猴病毒40转化的小鼠成纤维细胞部分纯化的质膜重构囊泡对氨基异丁酸进行钠刺激的主动转运。
Sodium-stimulated active transport of aminoisobutyric acid by reconstituted vesicles from partially purified plasma membranes of mouse fibroblasts transformed by simian virus 40.
作者信息
Nishino H, Tillotson L G, Schiller R M, Inui K I, Isselbacher K J
出版信息
Proc Natl Acad Sci U S A. 1978 Aug;75(8):3856-8. doi: 10.1073/pnas.75.8.3856.
Plasma membrane fractions isolated from mouse fibroblast BALB/c 3T3 cells transformed by simian virus 40 were partially purified by treatment with dimethylmaleic anhydride followed by extraction with 2% cholate. The extracted proteins were combined with exogenous phospholipids and eluted through a Sephadex G50 column. Reconstituted vesicles thus obtained were shown to possess the ability of Na+-stimulated transport of alpha-aminoisobutyric acid. The simultaneous addition of NaSCN and alpha-aminoisobutyric acid to these vesicles produced a transient accumulation above the equilibrium level (overshoot, active transport). The Na+-stimulated transport of alpha-aminoisobutyric acid was sensitive to the accompanying anion and to the temperature of incubation. The results demonstrate that partially purified membrane proteins of mouse fibroblast cells can be incorporated into the liposomes that have the characteristics of Na+-stimulated and electrochemically sensitive active transport of alpha-aminoisobutyric acid.
从经猿猴病毒40转化的小鼠成纤维细胞BALB/c 3T3细胞中分离得到的质膜组分，先用顺丁烯二酸酐处理，再用2%胆酸盐提取，进行部分纯化。将提取的蛋白质与外源性磷脂混合，通过葡聚糖G50柱洗脱。由此获得的重构囊泡显示出具有Na⁺刺激的α-氨基异丁酸转运能力。向这些囊泡中同时加入硫氰酸钠和α-氨基异丁酸会产生高于平衡水平的瞬时积累（过冲，主动转运）。α-氨基异丁酸的Na⁺刺激转运对伴随的阴离子和孵育温度敏感。结果表明，小鼠成纤维细胞的部分纯化膜蛋白可以掺入具有Na⁺刺激和α-氨基异丁酸电化学敏感主动转运特性的脂质体中。
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