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Single molecule observation of hard–soft-acid–base (HSAB) interaction in engineered Mycobacterium smegmatis porin A (MspA) nanopores

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Abstract

In the formation of coordination interactions between metal ions and amino acids in natural metalloproteins, the bound metal ion is critical either for the stabilization of the protein structure or as an enzyme co-factor. Though extremely small in size, metal ions, when bound to the restricted environment of an engineered biological nanopore, result in detectable perturbations during single channel recordings. All reported work of this kind was performed with engineered α-hemolysin nanopores and the observed events appear to be extremely small in amplitude (∼1–3 pA). We speculate that the cylindrical pore restriction of α-hemolysin may not be optimal for probing extremely small analytes. Mycobacterium smegmatis porin A (MspA), a conical shaped nanopore, was engineered to interact with Ca2+, Mn2+, Co2+, Ni2+, Zn2+, Pb2+ and Cd2+ and a systematically larger event amplitude (up to 10 pA) was observed. The measured rate constant suggests that the coordination of a single ion with an amino acid follows hard–soft-acid–base theory, which has never been systematically validated in the case of a single molecule. By adjusting the measurement pH from 6.8 to 8.0, the duration of a single ion binding event could be modified with a ∼46-fold time extension. The phenomena reported suggest MspA to be a superior engineering template for probing a variety of extremely small analytes, such as monatomic and polyatomic ions, small molecules or chemical intermediates, and the principle of hard–soft-acid–base interaction may be instructive in the pore design.

Graphical abstract: Single molecule observation of hard–soft-acid–base (HSAB) interaction in engineered Mycobacterium smegmatis porin A (MspA) nanopores

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Supplementary files

Article information


Submitted
17 Oct 2019
Accepted
03 Dec 2019
First published
10 Dec 2019

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2020,11, 879-887
Article type
Edge Article

Single molecule observation of hard–soft-acid–base (HSAB) interaction in engineered Mycobacterium smegmatis porin A (MspA) nanopores

S. Wang, J. Cao, W. Jia, W. Guo, S. Yan, Y. Wang, P. Zhang, H. Chen and S. Huang, Chem. Sci., 2020, 11, 879
DOI: 10.1039/C9SC05260G

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