Investigation of protein FTT1103 electroactivity using carbon and mercury electrodes. Surface-inhibition approach for disulfide oxidoreductases using silver amalgam powder

[Display omitted] •The complex electrochemical investigation of protein FTT1103 is described.•Hg-containing and carbon electrodes were used for analysis of the protein at nanomolar level.•Amalgam electrode was used in poorly water-soluble protein analysis for the first time.•Surface-inhibition appro...

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Bibliographic Details
Published in:	Analytica chimica acta Vol. 830; pp. 23 - 31
Main Authors:	Večerková, Renata, Hernychová, Lenka, Dobeš, Petr, Vrba, Jiří, Josypčuk, Bohdan, Bartošík, Martin, Vacek, Jan
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 09-06-2014
Subjects:	Adsorption Bacteria Carbon - chemistry Disulfide bond forming protein Disulfides Electrochemical analysis Electrochemical sensing Electrochemical Techniques Electrodes Membrane proteins Mercury - chemistry Models, Molecular Nanostructure Oxidoreductase Particle Size Powders - chemistry Protein Disulfide Reductase (Glutathione) - analysis Protein Disulfide Reductase (Glutathione) - antagonists & inhibitors Protein Disulfide Reductase (Glutathione) - metabolism Proteins Recombinant Proteins - analysis Recombinant Proteins - metabolism Silver - chemistry Silver - pharmacology Surface chemistry Surface inhibition Surface Properties Voltammetry SDS CPSA DTT Oxidoreductase HMDE Disulfide bond forming protein PAGE AgSA EDTA CHER Surface inhibition WE PGE Membrane proteins BSA SWV ACV AgSAE W Y Electrochemical sensing TRX TCEP
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Summary:	[Display omitted] •The complex electrochemical investigation of protein FTT1103 is described.•Hg-containing and carbon electrodes were used for analysis of the protein at nanomolar level.•Amalgam electrode was used in poorly water-soluble protein analysis for the first time.•Surface-inhibition approach presented could be used for other (not only DsbA) protein studies. 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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 ObjectType-Article-2 ObjectType-Feature-1
ISSN:	0003-2670 1873-4324
DOI:	10.1016/j.aca.2014.04.056