高等学校化学学报

高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (3): 628.doi: 10.7503/cjcu20120654

• 物理化学 • 上一篇    下一篇

分子对接和动力学模拟提高嗜热蛋白酶PhpI的活力

詹冬玲1,2, 高楠3, 韩葳葳1, 冯雁1   

  1. 1. 吉林大学分子酶学工程教育部重点实验室, 长春 130012;
    2. 吉林农业大学食品科学与工程学院, 长春 130118;
    3. 中国科学院长春应用化学研究所, 长春130022
  • 收稿日期:2012-07-10 出版日期:2013-03-10 发布日期:2013-02-18
  • 通讯作者: 韩葳葳,女,博士,副教授,主要从事计算生物学研究.E-mail:weiweihan@jlu.edu.cn;冯雁,女,博士,教授,主要从事嗜热酶的研究.E-mail:yfeng@jlu.edu.cn E-mail:weiweihan@jlu.edu.cn;yfeng@jlu.edu.cn
  • 基金资助:

    国家"九七三"计划项目(批准号:2012CB721003)和国家自然科学基金(批准号:31070638)资助.

Molecular Docking and Dynamics Simulation Improving Thermophilic Protease Activity of PhpI

ZHAN Dong-Ling1,2, GAO Nan3, HAN Wei-Wei1, FENG Yan1   

  1. 1. Key Laboratory for Molecular Enzymology and Engineering, Ministry of Education, Jilin University, Changchun 130012, China;
    2. College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China;
    3. Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China
  • Received:2012-07-10 Online:2013-03-10 Published:2013-02-18
  • Contact: Weiwei Han E-mail:weiweihan@jlu.edu.cn;yfeng@jlu.edu.cn

PDF (PC)

被引次数

19

摘要：

通过分子对接和动力学模拟对嗜热蛋白酶的分子进行改造, 确定蛋白酶PH1704(PhpI)定点突变残基, 并通过分子生物学实验进行验证. 突变体K43C的蛋白酶活力提高了5.8倍. 分子动力学模拟结果表明, 经过8 ns的动力学模拟后, K43C突变体二级结构由野生型的S2片层(F11-E12-D13)变成环状结构. E12和K43均是活性位点的重要残基, 这种变化将导致活性位点的柔性增强, 有利于催化反应的发生.

关键词: 嗜热蛋白酶PhpI, 分子对接, 动力学模拟, 定点突变

Abstract:

In this study, flexible docking approach was employed to dock the substrate into the active site of protease PH1704(PhpI), combining with crystal structure to determine the key enzyme, and study on site-mutation in theroy. All parameters were verified by molecular biology experiment. The protease activity of K43C was 5.8 times higher than that of WT. Molecular dynamics simulation reveals that after 8 ns dynamics simulations, the secondary structure of K43C mutant changes from the S2 sheet of WT(F11-E12-D13)to the loop. K43 and E12 are both the important active site residues. The change will enhance the flexibility of active site, and be helpful for catalytic reactions. The new structural and mechanistic insights obtained from molecular docking and dynamics simulation should be valuable for detailed researching of structures and mechanisms of the member of the DJ-1 superfamily.

Key words: Thermophilic protease PhpI, Molecular docking, Dynamics simulation, Site mutation

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