Glucose transport carrier activities in extensively washed human red cell ghosts.
Glucose transport carrier activities in extensively washed human red cell ghosts.
复制标题
广泛洗涤的人红细胞血影中的葡萄糖转运载体活性。
DOI：
10.1016/0005-2736(71)90059-9
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发表时间：
1971
期刊：
影响因子：
--
通讯作者：
D. Whaley
中科院分区：
文献类型：
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作者：
C. Jung;L. M. Carlson;D. Whaley
制备人红细胞血影，并在二价阳离子（Ca ~（2+）和Mg ~（2+））存在下进行广泛洗涤，并研究其残留葡萄糖转运载体活性。99.18%的血红蛋白原始细胞内容物被分离程序除去。这些血影的葡萄糖载体活性表现出以下特征。(1)与d-甘露醇的渗透性（24°时为5.2·10− 8 cm/sec）相比，d-葡萄糖（24°时为2.0·10− 5cm/sec）和2-脱氧-d-葡萄糖（24 °时为2.9·10−5cm/sec）的渗透性极高。(2)对各种糖的立体特异性; 2-脱氧-D-葡萄糖，D-葡萄糖，D-甘露糖和D-核糖的相对渗透率分别为1.5，1.0，0.35和0.020。(3)饱和度（24°时的福特葡萄糖，aKm为19.5 mM，Φ max为6.4 μ mol·cm−3·sec−2）。(4)对己烯雌酚的敏感性（24°时aKi为4.69 μM的竞争性抑制）和对二硝基氟苯的敏感性（不可逆抑制）。(5)通过添加第二种糖2-脱氧-d-葡萄糖，d-葡萄糖的瞬时上坡运动，一种逆转运现象。(6)对pH值变化相对不敏感，在5.2和9.0之间。(7)高度的温度依赖性，Kmat低温时为负Q10。(8)热失活的相对电阻率高达65°。总之，这项工作提供了（1）第一个证据表明，这种纯化的鬼保持完全功能的葡萄糖载体活性和（2）一个新的详细描述的性质的温度依赖性和热变性的载体系统在鬼。
Human red cell ghosts were prepared and extensively washed in the presence of divalent cations (Ca2+and Mg2+) and their residual glucose transport carrier activities were studied. 99.18 % of the original cellular contents of hemoglobin were removed by the isolation procedures. The glucose carrier activities of such ghosts exhibited the following characteristics. (1) An extremely high permeability tod-glucose (2.0·10−5cm/sec at 24°) and to 2-deoxy-d-glucose (2.9·10−5cm/sec at 24°), compared with the permeability tod-mannitol (5.2·10−8cm/sec at 24°). (2) Stereospecificity towards various sugars; relative rates of permeation by 2-deoxy-d-glucose,d-glucose,d-mannose andd-ribose were 1.5, 1.0, 0.35 and 0.020, respectively. (3) Saturability (ford-glucose at 24°, aKmof 19.5 mM and aΦmaxof 6.4 μmoles·cm−3·sec−2). (4) Sensitivity to stilbestrol (competitive inhibition with aKiof 4.69 μM at 24°) and to dinitrofluorobenzene (irreversible inhibition). (5) Transient uphill movements ofd-glucose by the addition of a second sugar, 2-deoxy-d-glucose, a counter-transport phenomenon. (6) Relative insensitivity to change in pH, between 5.2 and 9.0. (7) High degree of temperature dependency with a negativeQ10forKmat low temperatures. (8) A relative resistivity to heat inactivation up to 65°.In conclusion, this work provides (1) the first evidence that such purified ghosts maintain fully functional glucose carrier activity and (2) a new detailed description of the nature of the temperature dependence and heat denaturation of this carrier system in ghosts.