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用三硝基苯磺酸对从深红红螺菌和棕色固氮菌中分离出的胞内膜的脂质和蛋白质进行化学修饰。

Chemical modification by trinitrobenzenesulfonate of a lipid and proteins of intracytoplasmic membranes isolated from Chromatium vinosum and Azotobacter vinelandii.

作者信息

Shimada K, Murata N

出版信息

Biochim Biophys Acta. 1976 Dec 14;455(3):605-20. doi: 10.1016/0005-2736(76)90035-3.

Abstract
  1. The structure of intracytoplasmic membranes of a photosynthetic bacterium chromatium vinosum and a nitrogen-fixing bacterium Azotobacter vinelandii was studied by chemical modification of amino groups of phosphatidylethanolamine and proteins with trinitrobenzenesulfonate. 2. Almost all the constituents of intracytoplasmic membranes of C. vinosum were solubilized in a mixture of chloroform, methanol and trichloroacetic acid. One-third of proteins in the intracytoplasmic membranes of C. vinosum was found solubilized in a mixture of chloroform and methanol. By using a column chromatography with Sephadex LH-20 in organic solvents, the unmodified as well as the trinitrophenylated proteins and also the trinitrophenylated phosphatidylethanolamine were separated from the other colored substances. 3. In the chemical modification of the intracytoplasmic membrane preparations, 30% of phosphatidylethanolamine and 15% of protein amino groups in C. vinosum and 45% of phosphatidylethanolamine and 20% of protein amino groups in A. vinelandii were estimated to be exposed to the aqueous phase. In the single-layered liposomes composed of phosphatidylethanolamine and phosphatidylglycerol with a ratio of 2:1, 40% of phosphatidylethanolamine were estimated to be exposed to the aqueous phase.
摘要
  1. 利用三硝基苯磺酸对磷脂酰乙醇胺和蛋白质的氨基进行化学修饰,研究了光合细菌嗜酒色杆菌和固氮细菌维涅兰德固氮菌胞内膜的结构。2. 嗜酒色杆菌胞内膜的几乎所有成分都可溶解于氯仿、甲醇和三氯乙酸的混合液中。发现嗜酒色杆菌胞内膜中三分之一的蛋白质可溶解于氯仿和甲醇的混合液中。通过在有机溶剂中使用葡聚糖凝胶LH - 20柱色谱法,将未修饰的以及三硝基苯基化的蛋白质和三硝基苯基化的磷脂酰乙醇胺与其他有色物质分离。3. 在胞内膜制剂的化学修饰中,估计嗜酒色杆菌中30%的磷脂酰乙醇胺和15%的蛋白质氨基以及维涅兰德固氮菌中45%的磷脂酰乙醇胺和20%的蛋白质氨基暴露于水相。在由磷脂酰乙醇胺和磷脂酰甘油按2:1比例组成的单层脂质体中,估计40%的磷脂酰乙醇胺暴露于水相。

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