Investigation of protein FTT1103 electroactivity using carbon and mercury electrodes. Surface-inhibition approach for disulfide oxidoreductases using silver amalgam powder
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
24856508
DOI
10.1016/j.aca.2014.04.056
PII: S0003-2670(14)00510-8
Knihovny.cz E-resources
- Keywords
- Disulfide bond forming protein, Electrochemical sensing, Membrane proteins, Oxidoreductase, Surface inhibition,
- MeSH
- Adsorption MeSH
- Electrochemical Techniques * MeSH
- Electrodes MeSH
- Models, Molecular MeSH
- Surface Properties MeSH
- Powders chemistry MeSH
- Protein Disulfide Reductase (Glutathione) analysis antagonists & inhibitors metabolism MeSH
- Recombinant Proteins analysis metabolism MeSH
- Mercury chemistry MeSH
- Silver chemistry pharmacology MeSH
- Carbon chemistry MeSH
- Particle Size MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Powders MeSH
- Protein Disulfide Reductase (Glutathione) MeSH
- Recombinant Proteins MeSH
- Mercury MeSH
- Silver MeSH
- Carbon MeSH
Recently, it was shown that electrochemical methods can be used for analysis of poorly water-soluble proteins and for study of their structural changes and intermolecular (protein-ligand) interactions. In this study, we focused on complex electrochemical investigation of recombinant protein FTT1103, a disulfide oxidoreductase with structural similarity to well described DsbA proteins. This thioredoxin-like periplasmic lipoprotein plays an important role in virulence of bacteria Francisella tularensis. For electrochemical analyses, adsorptive transfer (ex situ) square-wave voltammetry with pyrolytic graphite electrode, and alternating-current voltammetry and constant-current chronopotentiometric stripping analysis with mercury electrodes, including silver solid amalgam electrode (AgSAE) were used. AgSAE was used in poorly water-soluble protein analysis for the first time. In addition to basic redox, electrocatalytic and adsorption/desorption characterization of FTT1103, electrochemical methods were also used for sensitive determination of the protein at nanomolar level and study of its interaction with surface of AgSA microparticles. Proposed electrochemical protocol and AgSA surface-inhibition approach presented here could be used in future for biochemical studies focused on proteins associated with membranes as well as on those with disulfide oxidoreductase activity.
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