Control of Solubilized State and Estimation of Molecular Organization of Membrane Proteins by the Use of Analogues of Sodium Dodecyl Sulfate

使用十二烷基硫酸钠类似物控制溶解状态并估计膜蛋白的分子组织

基本信息

批准号：
60470157
负责人：
TAKAGI Toshio
金额：
$ 3.14万
依托单位：
Institute for Protein Research
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60470157/
关键词：
電気泳動生体膜蛋白質可溶化界面活性剤電気詠動

项目摘要

In the field of biochemical research,an anionic surfactant,sodium dodecyl sulfate(SDS),is routinely used.We have focused our attention on the potentiality of analogues of SDS of which general structure can be described as follows:n-C_<12>H_<25>(OCH_2CH_2)OSO^<3->X^+.It was found that:(1)among the analogues having their cations replaced by Li,Tris,triethanolammine and triisopropanolammine,the first was stronger even than SDS as protein denaturant,and the others were far less destructive than SDS as protein denaturant,and the others were far less destructive than SDSbecoming milder on the sequence mentioned above;(2)the series of surfactant could be used to determine the hierarchy of organization of molecular assemblies of proteins especially membrane proteins;(3)a typical example of the successful application of the series is the separation of the products of disassembly of thylakoid chlorophyllproteins in which different surfactant of the series giving different electrophoretic patterns;(4)analogues with m=2to8 were compared with to reveal that only those with m equal to or larger than 6 have high affinity to membrane proteins and thus allows efficient separation of mambrane prteins in the presence of such a surfactant;(5)a readily available surfactant similar to those described above with distribution with respect to both alkyl chain length (number of carbon atoms around 13) and oxyethylene chain length(around m =8)was found to be an efficient additive in polyacrylamide gel elctrophoresis for separation of the subunits of (Na,K)-ATPase without causing denaturation.

在生化研究领域，通常使用一种阴离子表面活性剂十二烷基硫酸钠（SDS）。我们将注意力集中在SDS类似物的潜力上，可以将一般结构描述如下：N-C_ <12> H_ <25>（OCH_2CH_2）OSO^<3-> X^+。发现：（1）在被LI，Tris，Tris，Triethanolammine和Triisopanolammine取代的类似物中，第一个甚至比SDS更强大变性剂，其他变性剂的破坏性远不如SDS作为蛋白质变性剂，而其他蛋白质变性剂的破坏性远不及上述序列的SDSBECOMINGINGIS;（2）一系列表面活性剂可用于确定分子组装组织的层次结构蛋白质特别是膜蛋白的蛋白质；（3）该系列成功应用成功应用的一个典型例子是分离囊样叶绿素叶绿素蛋白的产物，其中该系列的不同表面活性剂提供不同的电泳模式；（4）M = 2to88的类似物与比较的是，只有M等于或大于6的人对膜蛋白具有很高的亲和力，因此可以在存在这种表面活性剂的情况下有效地分离Mambrane Prteins；（5）与上述相似的表面活性剂相似的表面活性剂在分布相对于两个烷基链长度（含13个碳原子的数量）和氧乙烯链长度（M = 8左右）的分布中，在聚丙烯酰胺凝胶elctrophoresis中是有效的添加剂，用于分离（Na，K）的亚基 - ATPase而不会引起变性。