Aerolysin单分子界面的构建及高选择性单分子检测

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

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.

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.