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Stereospecific hexose transport by membrane vesicles from mouse fibroblasts: Membrane vesicles retain increased hexose transport associated with viral transformation

机译：小鼠成纤维细胞膜囊泡的立体特异性己糖转运：膜囊泡保留与病毒转化有关的增加的己糖转运

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Membrane vesicles isolated from nontransformed BALB/c 3T3 mouse fibroblasts (3T3) and from those cells transformed by simian virus 40 (SV3T3) displayed carrier-mediated and stereospecific uptake of hexose as measured by the difference between D-[14C]glucose or its analogues and L-[3H]glucose uptake. Stereospecific uptake appeared to be linear for 5 sec and reached a maximum at 5-10 min. Stereospecific D-[14C]glucose uptake, osmotically sensitive and temperature dependent, was inhibited by unlabeled D-glucose or its analogues and was stimulated by the countertransport of accumulated unlabeled D-glucose. As with whole cells, the initial rate of stereospecific uptake by SV3T3 membrane vesicles was approximately 2.5-fold greater than that by 3T3 vesicles. Efflux of preloaded D-[14C]glucose was also faster from SV3T3 than from 3T3 membrane vesicles. The Km value was 5 mM for both the 3T3 and the SV3T3 membrane vesicles, but the Vmax values were 36 and 86 nmol/mg of protein per min, respectively, suggesting an increase in the number or availability of hexose carriers in transformed cell membranes. Cytochalasin B competitively inhibited stereospecific hexose uptake in both types of membrane vesicles. The binding of cytochalasin B to the SV3T3 membrane vesicles was significantly greater than that to 3T3 vesicles. Thus, the membrane vesicles retained many of the features of the altered hexose transport observed in whole cells in association with viral transformation.
机译：从未转化的BALB / c 3T3小鼠成纤维细胞（3T3）和猿猴病毒40（SV3T3）转化的细胞中分离出的膜囊泡显示了己糖的载体介导和立体定向摄取，通过D- [14C]葡萄糖或其类似物之间的差异进行测量和L- [3H]葡萄糖摄取。立体定向摄取在5秒钟内呈线性变化，并在5-10分钟达到最大值。渗透敏感和温度依赖性的立体特异性D- [14C]葡萄糖摄取受到未标记的D-葡萄糖或其类似物的抑制，并受到积累的未标记D-葡萄糖的逆向运输的刺激。与全细胞一样，SV3T3膜囊泡的立体定向摄取的初始速率比3T3囊泡的立体定向摄取的初始速率高约2.5倍。 SV3T3的预载D- [14C]葡萄糖流出速度也快于3T3膜囊泡的流出速度。 3T3和SV3T3膜囊泡的Km值均为5 mM，但Vmax值分别为每分钟36和86 nmol / mg蛋白质，表明转化细胞膜中己糖载体的数量或可用性增加。细胞松弛素B在两种类型的膜囊泡中均竞争性抑制立体特异性己糖摄取。细胞松弛素B与SV3T3膜囊泡的结合显着大于与3T3膜囊泡的结合。因此，膜囊泡保留了在全细胞中观察到的与病毒转化相关的己糖转运改变的许多特征。

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