Highly Selective Separation of Rhodopsin from Bovine Rod Outer Segment Membranes Using Combination of Divalent Cation and Alkyl(thio)glucoside
Highly Selective Separation of Rhodopsin from Bovine Rod Outer Segment Membranes Using Combination of Divalent Cation and Alkyl(thio)glucoside
复制标题
利用二价阳离子和烷基（硫代）葡萄糖苷的组合从牛视杆外节膜中高度选择性分离视紫红质
DOI：
10.1111/j.1751-1097.1998.tb09084.x
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发表时间：
1998
影响因子：
3.3
通讯作者：
T. Kouyama
中科院分区：
文献类型：
--
作者：
T. Okada;Kazuki Takeda;T. Kouyama
研究了一系列中性洗涤剂对牛骨棒外节膜胶团化过程的影响。研究发现，当具有适当的亲油-亲脂平衡的烷基（硫代）葡糖苷（例如辛基硫代葡糖苷）与二价阳离子组合使用时，视紫红质以特定的去污剂-膜比从ROS膜中选择性提取。这允许通过一步增溶来显著纯化视紫红质，因为残留的膜在二价阳离子存在下严重聚集，因此容易通过低速离心沉淀。上清液的吸收光谱可再现地显示出A280/A500值为1.6，这是一个很好的值，很少能通过色谱纯化获得。纯化的程度也取决于溶解溶液中包含的二价阳离子的类型; IIB系列阳离子（Zn 2+和Cd 2+）与ROS膜的特异性结合被认为在溶解过程中起重要作用。本结果代表了从高度聚集的膜中选择性溶解特定膜蛋白的独特实例。
The micellization process of bovine rod outer segment (ROS) membranes is investigated utilizing a series of neutral detergents. It is found that when alkyl(thio)glucosides with an appropriate hydrophilic–lipophilic balance (e.g. octylthioglucoside) are used in combination with a divalent cation, rhodopsin is selectively extracted from ROS membranes at a specific detergent‐to‐membrane ratio. This allows remarkable purification of rhodopsin by a single‐step solubilization, because the residual membranes are heavily aggregated in the presence of divalent cation and are therefore easily sedimented by low‐speed centrifugation. The absorption spectrum of the supernatant reproducibly exhibits an A280/A500 value of 1.6, an excellent value that could rarely be obtained by chromatographic purification. The degree of purification also depends on the type of divalent cation included in the solubilization solution; specific binding of IIB‐series cations (Zn2+ and Cd2+) to ROS membranes is suggested to play an important role in the solubilization process. The present result represents a unique example of selective solubilization of a specific membrane protein from highly aggregated membranes.