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

Večerková, Renata; Hernychová, Lenka; Dobeš, Petr; Vrba, Jiřı́; Josypčuk, Bohdan; Bartošík, Martin; Vacek, Jan

doi:10.1016/j.aca.2014.04.056

Cited by 12 publications

(18 citation statements)

References 40 publications

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“…As expected, complementation with the LVS dsbA CXXS active‐site mutant did not restore copper resistance (Fig. A) because CXXC is required for F. tularensis oxidoreductase activity (Schmidt et al ., ; Vecerkova et al ., ). In reference to the second DsbA ortholog, FTL_1550, described above; ΔFTL_1550 was found to be as copper resistant as WT, indicating that FTL_1550 does not affect F. tularensis isomerase activity (http://onlinelibrary.wiley.com/doi/10.1111/mmi.12808/suppinfo).…”

Section: Resultsmentioning

confidence: 97%

Identification of disulfide bond isomerase substrates reveals bacterial virulence factors

Ren

Champion

Huntley

2014

Molecular Microbiology

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Summary Bacterial pathogens are exposed to toxic molecules inside the host and require efficient systems to form and maintain correct disulfide bonds for protein stability and function. The intracellular pathogen Francisella tularensis encodes a disulfide bond formation protein ortholog, DsbA, which previously was reported to be required for infection of macrophages and mice. However, the molecular mechanisms by which F. tularensis DsbA contributes to virulence are unknown. Here, we demonstrate that F. tularensis DsbA is a bifunctional protein that oxidizes and, more importantly, isomerizes complex disulfide connectivity in substrates. A single amino acid in the conserved cis-proline loop of the DsbA thioredoxin domain was shown to modulate both isomerase activity and F. tularensis virulence. Trapping experiments in F. tularensis identified over 50 F. tularensis DsbA substrates, including outer membrane proteins, virulence factors, and many hypothetical proteins. Six of these hypothetical proteins were randomly selected and deleted, revealing two novel proteins, FTL_1548 and FTL_1709, which are required for F. tularensis virulence. We propose that the extreme virulence of F. tularensis is partially due to the bifunctional nature of DsbA, that many of the newly-identified substrates are required for virulence, and that the development of future DsbA inhibitors could have broad anti-bacterial implications.

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Section: Resultsmentioning

confidence: 97%

Identification of disulfide bond isomerase substrates reveals bacterial virulence factors

Ren

Champion

Huntley

2014

Molecular Microbiology

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“…The results of the NKA analysis 411 show that CPS analysis can be used not only for studies of structural changes and intermolecular interactions in water-soluble proteins at bare 155 , 230 , 232 , 233 , 238 , 254 , 256 , 322 and thiol-modified electrodes, 105 , 228 , 238 , 239 but also for the investigation of poorly soluble proteins. 410 , 411 , 423 …”

Section: Label-free Protein Analysis In Biomedicinementioning

confidence: 99%

Electrochemistry of Nonconjugated Proteins and Glycoproteins. Toward Sensors for Biomedicine and Glycomics

Paleček

Tkáč²,

Bartošík³

et al. 2015

Chem. Rev.

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287

276

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“…In addition to this, the methodology proposed could be the starting point for the further development of novel techniques for protein glycation detection, especially electrochemical sensors and versatile amalgam electrode-based sensing platforms. 35,36 ■ ASSOCIATED CONTENT * S Supporting Information [Scheme S1, graphical representation of sample handling and analytical methods used for detecting protein glycation; Figure S1, details on early glycation product detection and quantitative analysis of BSA; Table S1, overview of methods for BSA glycation analysis; Table S2, number of total and surfaceexposed Cys, Lys, Arg, and His residues in BSA molecule. This material is available free of charge via the Internet at http:// pubs.acs.org.…”

Section: ■ Conclusionmentioning

confidence: 99%

Electrocatalytic Assay for Monitoring Methylglyoxal-Mediated Protein Glycation

et al. 2015

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Protein glycation is a complex process that plays an important role in diabetes mellitus, aging, and the regulation of protein function in general. As a result, current methodological research on proteins is focused on the development of novel approaches for investigating glycation and the possibility of monitoring its modulation and selective inhibition. In this paper, a first sensing strategy for protein glycation is proposed, based on protein electroactivity measurement. Concretely, the label-free method proposed is based on the application of a constant-current chronopotentiometric stripping (CPS) analysis at Hg-containing electrodes. The glycation process was monitored as the decrease in the electrocatalytic protein signal, peak H, observed at highly negative potentials at around -1.8 V (vs Ag/AgCl3 M KCl), which was previously ascribed to a catalytic hydrogen evolution reaction (CHER). Using this method, a model protein bovine serum albumin was investigated over 3 days of incubation with the glycation agent methylglyoxal in the absence or presence of the glycation inhibitor aminoguanidine (pimagedine). The electrochemical methodology presented here could open up new possibilities in research on protein glycation and oxidative modification. The methodology developed also provides a new option for the analysis of protein intermolecular interactions using electrochemical sensors, which was demonstrated by the application of a silver solid amalgam electrode (AgSAE) for monitoring the glycation process in samples of bovine serum albumin, human serum albumin, and lysozyme.

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

Cited by 12 publications

References 40 publications

Identification of disulfide bond isomerase substrates reveals bacterial virulence factors

Identification of disulfide bond isomerase substrates reveals bacterial virulence factors

Electrochemistry of Nonconjugated Proteins and Glycoproteins. Toward Sensors for Biomedicine and Glycomics

Electrocatalytic Assay for Monitoring Methylglyoxal-Mediated Protein Glycation

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