高等学校化学学报 ›› 2019, Vol. 40 ›› Issue (9): 1825.doi: 10.7503/cjcu20190364

• 分析化学 • 上一篇    下一篇

Aerolysin单分子界面的构建及高选择性单分子检测

武雪原1,应佚伦1,2,龙亿涛1,2,*()   

  1. 1. 华东理工大学化学与分子工程学院, 上海 200237
    2. 南京大学化学化工学院, 生命分析化学重点实验室, 南京 210023
  • 收稿日期:2019-07-01 出版日期:2019-09-10 发布日期:2019-09-09
  • 通讯作者: 龙亿涛 E-mail:ytlong@ecust.edu.cn;yitaolong@nju.edu.cn
  • 基金资助:
    国家自然科学基金(21834001, 61871183)

Construction and High Selectivity of Aerolysin Single Molecular Interface for Single Molecule Analysis

WU Xueyuan1,YING Yilun1,2,LONG Yitao1,2,*()   

  1. 1. School of Chemistry and Molecular Engineering, East China University of Science and Technology,Shanghai 200237, China
    2. State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering,Nanjing University, Nanjing 210023, China
  • Received:2019-07-01 Online:2019-09-10 Published:2019-09-09
  • Contact: LONG Yitao E-mail:ytlong@ecust.edu.cn;yitaolong@nju.edu.cn
  • Supported by:
    Supported by the National Natural Science Foundation of China(21834001, 61871183)

摘要:

提出了一种基于Aerolysin膜蛋白质分子构建单分子界面的方法, 运用蛋白质工程技术对单分子界面进行定点修饰, 所建立方法灵活、 可控且重复性好. 采用Poly(dA)4为探针分子对修饰后的单分子界面进行了表征, 结果表明, 在孔口处的Arginine修饰影响了寡聚核苷酸的选择性. 为进一步理解Aerolysin单分子界面及合理设计功能性单分子界面提供了参考.

关键词: 气单胞菌溶素, 单分子界面, 纳米孔道, 单分子分析

Abstract:

As an new conceptual single molecular interface, the biomolecular nanopore can accommodate only a single molecule and contains abundant amino acid residues in the lumen, providing an appropriate confinement for single molecule detection. Herein, a generalized method is proposed for the construct of single molecular interface based on membrane protein. The method is flexible, controllable and repeatable. The characterization of D209R mutation with Poly(dA)4 illustrate that the selectivity of aerolysin single molecular interface is related the electrostatic properties at the entrance of nanopore, which deepens our understanding of aerolysin single molecular interface, providing a paradigm for rational design of functional single molecular interface.

Key words: Aerolysin, Single molecular interface, Nanopores, Single molecule analysis

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