Citation: ZHUO Sha, WANG Ying, KANG Xiao-Feng. Engineered Protein Nanopore for Real-time Monitoring Single-molecule Reaction between Cadmium Ion and Glutathione[J]. Chinese Journal of Analytical Chemistry, ;2017, 45(8): 1172-1179. doi: 10.11895/j.issn.0253-3820.170109 shu

Engineered Protein Nanopore for Real-time Monitoring Single-molecule Reaction between Cadmium Ion and Glutathione

College of Chemistry and Materials Science, Northwest University, Xi'an 710127, China

Received Date: 23 February 2017
Revised Date: 27 May 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos.21175105,21375104,21327806).

The chelating reaction between glutathione peptides and divalent cadmium ion was used as a typical model for investigating the coordination chemistry of SH-containing peptides and heavy metal ions, which was essential to understand the mechanism of intracellular cadmium detoxification. Here, the mutant (CM113R)₇ αHL protein nanopore equipped with a first synthesized per-6-quaternary ammonium-β-cyclodextrin (p-QABCD) was used as nanoreactor and detector to investigate the single-molecule reaction of GSH molecules and Cd²⁺ ions. Different reaction pathways, intermediates, and products could be recognized. Cd²⁺-GSH reaction was highly dependent on the solution pH. The Cd(GSH)₂ was formed at pH 7.4, while the Cd(GSH)₂ and Cd₂(GSH)₂ were formed at pH 9.0. Cd₂(GSH)₂ was formed by two possible pathways:(1) A Cd²⁺ ion primarily coordinated with the thiol group of two GSH molecules to form Cd(GSH)₂, and then the second Cd²⁺ ion quickly incorporated with the deprotonated amino group of Cd(GSH)₂ to form Cd₂(GSH)₂; (2) Two Cd²⁺ ions separately coordinated with the thiol and deprotonated amino group of one GSH molecule to form Cd₂(GSH)₁, and the second GSH molecule quickly bounded to Cd²⁺ ions to form Cd₂(GSH)₂. This work studied the chelating reaction between metal ions and bioactive glutathione at single-molecule level without labeling and chemical modification, and would be important to further understand intracellular mechanisms of detoxification of heavy metals and greatly expand the research field of nanopore single-molecule technique.
- Nanopore,
- Single-molecule detection,
- Per-6-quaternary ammonium-β-cyclodextrin,
- Glutathione,
- Cadmium ion
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