Isolation of the sodium-dependent d-glucose transport protein from brush-border membranes.
Isolation of the sodium-dependent d-glucose transport protein from brush-border membranes.
复制标题
从刷状缘膜中分离钠依赖性 d-葡萄糖转运蛋白。
DOI：
10.1016/0005-2736(83)90144-x
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
H. Preiser
中科院分区：
文献类型：
--
作者：
P. Malathi;H. Preiser
将兔肾刷状缘膜囊泡暴露于细菌蛋白水解酶中，分解出大量的外向蛋白。在酶处理的囊泡中，钠离子依赖的葡萄糖转运保持不变。酶处理的膜用脱氧胆酸增溶，脱氧胆酸提取的蛋白质通过ConA-Sepharose柱进一步分离。钠依赖的葡萄糖活性被发现存在于含有MR≅165000的单一蛋白质带的一部分中，这显然是MR≅85000的二聚体。当重组并测试转运时，该蛋白表现出钠离子依赖、立体特异性和根癌苷抑制的葡萄糖转运。运输活动完全恢复，比活动增加20倍。从兔小肠刷状缘膜和其他几种动物的肾脏中也获得了类似的分离物。
Rabbit kidney brush-border membrane vesicles were exposed to bacterial protease which cleaves off a large number of externally oriented proteins. Na+-dependentd-glucose transport is left intact in the protease-treated vesicles. The protease-treated membrane was solubilized with deoxycholate and the deoxycholate-extracted proteins were further resolved by passage through Con A-Sepharose columns. Sodium-dependentd-glucose activity was found to reside in a fraction containing a single protein band ofMr≅ 165000 which is apparently a dimer ofMr≅ 85 000. When reconstituted and tested for transport, this protein showed Na+-dependent, stereo-specific and phlorizin-inhibitable glucose transport. Transport activity is completely recovered and is 20-fold increased in specific activity. A similar isolate was obtained from rabbit small intestinal brush-border membranes and kidneys from several other species of animals.