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Citation: DUAN Jing, ZHOU Sha, YAO Fu-Jun, ZHANG Ya-Ni, KANG Xiao-Feng. A Single-Molecule Mycobacterium Smegmatis Porin A Protein Nanopore Sensor for Host-guest Chemistry[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(12): 1801-1807. doi: 10.11895/j.issn.0253-3820.160344 shu

A Single-Molecule Mycobacterium Smegmatis Porin A Protein Nanopore Sensor for Host-guest Chemistry

1.
College of Chemistry and Materials Science, Northwest University, Xi'an 710127, China;
2.
College of Life Science, Northwest University, Xi'an 710027, China

Received Date: 30 April 2016
Revised Date: 27 September 2016

Fund Project: This work was supported by the National Natural Foundation of China (Nos. 21175105, 21375104, 21327806) and the Ph.D. Programs Foundation of Ministry of Education of China (No.20126101110015).

A mycobacterium smegmatis porin A (MspA) protein nanopore was prepared by E.coil prokaryotic expression system. The extraction experiments of MspA nanopore using surfactants showed that an extraction agent concentration of 0.5%, a temperature of 90℃ and an extraction time of 30 min were the optimum extraction conditions. The interactions of MspA nanopore with a variety of cyclodextrins were examined at single-molecule level. The interaction between MspA and Per-6-amino-β-cyclodextrin (am₇-β-CD) was stronger and consistent with blocking platform. Its retention time in the MspA nanopore decreased with the increasing voltage, whereas the blocking current was almost not affected by voltage. So the am₇-β-CD was selected as a non-covalent adapter and lodged within single MspA nanopore to study the single-molecule host-guest chemistry. After adding am₇-β-CD to form a first blocked platform, a small concentration of 50 μmol/L for the amantadine hydrochloride showed a second blocked platform and its residence time decreased with the increasing voltage. This new protein nanopore sensor can be used as a single-molecule detector and identifier for small organic molecules, which will greatly broaden the application range of MspA nanopore.
- Mycobacterium smegmatis porin A,
- Nanopore,
- Single-molecule detection,
- Cyclodextrin,
- 1-Amantadine hydrochloride
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