Parnes J R, Garvey T Q, Isselbacher K J
J Cell Physiol. 1976 Dec;89(4):789-94. doi: 10.1002/jcp.1040890444.
Mixed membrane vesicle populations composed of plasma membrane and endoplasmic reticulum were prepared from Balb/c 3T3 and simian virus 40-transformed Balb/c 3T3 mouse fibroblasts. The initial rates of uptake of L-leucine and alpha-aminoisobutyric acid by these vesicles were stimulated by a NaCl gradient (external greater than internal). Cation specificity for stimulation of L-leucine uptake was Na+ greater than Li+ greater than K+. NaSCN was as effective as NaCl. Stimulation of uptake of both amino acids by a NaCl gradient was twice as great in vesicles from transformed as compared to non-transformed cells. The NaCl gradient produced transient accumulation of both L-leucine and alpha-aminoisobutyric acid to twice the equilibrium level in vesicles from transformed cells. No such "overshoot" was observed in vesicles from nontransformed cells. In vesicles from the contact-inhibitable Balb/c 3T3 cells, transport of alpha-aminoisobutyric acid, but non L-leucine, exhibited a density-dependent decrease in Na+ gradient induced stimulation, from 248% for sub-confluent to 109% with confluent cells. No density-related changes in uptake were noted with vesicles from the transformed cells. These studies suggest that variation in amino acid uptake associated with viral transformation may be related, at least in part, to alterations in Na+ permeability of the surface membrane.
从Balb/c 3T3细胞和猿猴病毒40转化的Balb/c 3T3小鼠成纤维细胞中制备了由质膜和内质网组成的混合膜泡群体。这些膜泡对L-亮氨酸和α-氨基异丁酸的初始摄取速率受到NaCl梯度(外部大于内部)的刺激。刺激L-亮氨酸摄取的阳离子特异性为Na+>Li+>K+。硫氰酸钠与氯化钠的效果相同。与未转化细胞相比,来自转化细胞的膜泡中,NaCl梯度对两种氨基酸摄取的刺激作用是其两倍。NaCl梯度使来自转化细胞的膜泡中L-亮氨酸和α-氨基异丁酸都短暂积累至平衡水平的两倍。在未转化细胞的膜泡中未观察到这种“过冲”现象。在接触抑制性Balb/c 3T3细胞的膜泡中,α-氨基异丁酸而非L-亮氨酸的转运在Na+梯度诱导的刺激中表现出密度依赖性降低,从亚汇合细胞的248%降至汇合细胞的109%。来自转化细胞的膜泡未观察到与密度相关的摄取变化。这些研究表明,与病毒转化相关的氨基酸摄取变化可能至少部分与表面膜Na+通透性的改变有关。