| 细菌视紫红质 E204Q 和 I119T/T121S/A126T突变体的构建及其功能研究 |
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| 引用本文： | 宋景娇,钟 声,张 悦,曹军卫,胡坤生.细菌视紫红质 E204Q 和 I119T/T121S/A126T突变体的构建及其功能研究[J].生物化学与生物物理进展,2005,32(6):529-534. |
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| 作者姓名： | 宋景娇 钟 声 张 悦 曹军卫 胡坤生 |
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| 作者单位： | 1. 武汉大学生命科学学院,武汉,430072
2. 中国科学院生物物理研究所,北京,100101 |
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| 基金项目： | 中国科学院知识创新工程重大项目(KJCX1-SW07)和国家自然科学基金资助项目(30170235). |
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| 摘 要： | 应用定点突变的方法获得了细菌视紫红质 (bacteriorhodopsin ， BR) 的单突变体 BRE204Q和三突变体 BRI119T/T121S/A126T. 通过功能研究发现， BR 蛋白的单突变体 BRE204Q的 M 态寿命为 7.10 ms ，三突变体 BRI119T/T121S/A126T的 M 态寿命为 8.23 ms ，均较野生型 BR 蛋白 (6.23ms) 有所延长，三突变体表现得更为显著，其 M 态延长时间可超出野生型的 32％. 同时单突变体 BRE204Q和三突变体 BRI119T/T121S/A126T的质子泵功能也均有改变，都比野生型 BR 蛋白有所下降，其中三突变体 BRI119T/T121S/A126T下降得更为明显.
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| 关 键 词： | 细菌视紫红质，突变体， M 态中间体，质子泵功能 |
Construction of Bacteriorhodopsin Mutant E204Q and I119T/T121S/A126T of PM Protein and Their Functional Research |
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| SONG Jing-Jiao,ZHONG Sheng,ZHANG Yue,CAO Jun-Wei and HU Kun-Sheng.Construction of Bacteriorhodopsin Mutant E204Q and I119T/T121S/A126T of PM Protein and Their Functional Research[J].Progress In Biochemistry and Biophysics,2005,32(6):529-534. |
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| Authors: | SONG Jing-Jiao ZHONG Sheng ZHANG Yue CAO Jun-Wei and HU Kun-Sheng |
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| Institution: | Department of Microbiology, Wuhan University, Wuhan 430072, China;Institute of Biophysis, The Chinese Academy of Sciences, Beijing 100101, China;Institute of Biophysis, The Chinese Academy of Sciences, Beijing 100101, China;Department of Microbiology, Wuhan University, Wuhan 430072, China;Institute of Biophysis, The Chinese Academy of Sciences, Beijing 100101, China |
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| Abstract: | The single mutant BRE204Q and tri-mutant BRI119T/T121S/A126T of bacteriorhodpsin were constructed by directed-mutation method. The M412 Flash-dynamics spectrum indicates that the life of M intermediate of the single mutant BRE204Q is 7.10 ms and the life of M intermediate of the tri-mutant BRI119T/T121S/A126T is 8.23 ms, whose lives of M intermediate are longer than that of the wild-type BR with life of the M intermediate 6.23 ms. The result shows that the tri-mutant has the longest M intermediate life which exceeds wild-type BR by thirty-two percent. The Proton-pump functional research of the two mutants illustrates that their proton-pump functions decresae compare with that of wild-type BR respectively and the proton-pump function of the tri-mutant BRI119T/T121S/A126T is the weakest one. |
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| Keywords: | bacteriorhodopsin mutant M intermediate proton-pump |
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