Detection of Abasic Sites in DNA by Electrochemical, Immunoelectrochemical and Acoustic Methods Using OsO<sub>4</sub>, 2,2′‐bipyridine as a Probe for Unpaired Thymine Residues

Bartošík, Martin; Gajdoš, Viktor; Kostečka, Pavel; Fojta, Miroslav; Paleček, Emil; Волков, Е. М.; Oretskaya, Tatiana S.; Hianik, Tibor

doi:10.1002/elan.200804380

Cited by 17 publications

(20 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2D) within dsDNA molecules, which are readily modified, leaving pyrimidine residues in the double helical parts intact. Increased reactivity of thymines [36,38] or 5-methylcytosines (5-mC) [21,22,27] was observed also within single nucleotide insertions (Fig. 2F), apurinic sites (Fig.…”

Section: Probing Dna Structure With Os(viii)lmentioning

confidence: 88%

“…The monoclonal antibody [63] was applied [65] in an enzyme-linked electrochemical assay of DNA hybridization using Os(VIII),bipy as immunomarker of the target DNA. The same antibody has recently been applied, in connection with an acoustic sensor, to detect osmium modification of unpaired thymine residues in DNA duplexes containing apurinic lesions [38].…”

Section: Detection Of Pyrimidine-osl Adducts In Dnamentioning

confidence: 99%

“…Using basal-plane pyrolytic graphite electrodes and the AdTS voltammetric procedure, it has been possible to study reactivity of various nucleic acid samples towards the Os(VIII),L (see examples of native and denatured DNA, as well as various homopolydeoxynuclotides, in Fig. 3C) [7], to detect enzymatic DNA cleavage and/or damage [37], to monitor unpaired thymines within abasic lesions [38], etc. Other applications include DNA hybridization assays with osmium-labeled targets [55] or signaling probes [17,56] (for more details see section 5).…”

Section: Carbon Electrodesmentioning

confidence: 99%

See 2 more Smart Citations

Osmium Tetroxide Complexes as Versatile Tools for Structure Probing and Electrochemical Analysis of Biopolymers

Fojta¹,

Kostečka²,

Pivoňková³

et al. 2011

CAC

Self Cite

View full text Add to dashboard Cite

Osmium tetroxide complexes with nitrogenous ligands and analogous complexes of six-valent osmium proved excellent tools for selective labeling of biopolymers (nucleic acids, proteins and polysaccharides). Reactions of these species with target moieties within the biopolymer molecules (pyrimidine nucleobases, tryptophan residues or sugar moieties) are facile at physiological conditions and are in general structure-selective, allowing their application in DNA and protein structure probing. The modification products can be detected by a variety of widely accessible analytical techniques, including biochemical (enzymatic) approaches, immunoassays, chemical DNA sequencing, spectrophotometry and electrochemistry. Particularly the electrochemical techniques are promising for utilization in biosensors and routine bioassays due to the possibility of highly sensitive and selective detection of the labeled biopolymers adducts based on distinct electrochemical properties of the introduced osmium moieties. Utilization of the osmium tags in probing DNA structural transitions, sensing of DNA hybridization, damage and DNA methylation, labeling of peptides and proteins, probing accessibility of tryptophan residues in proteins and their complexes, and labeling of sugar moieties, are reviewed.

show abstract

Section: Probing Dna Structure With Os(viii)lmentioning

confidence: 88%

Section: Detection Of Pyrimidine-osl Adducts In Dnamentioning

confidence: 99%

Section: Carbon Electrodesmentioning

confidence: 99%

See 1 more Smart Citation

Osmium Tetroxide Complexes as Versatile Tools for Structure Probing and Electrochemical Analysis of Biopolymers

Fojta¹,

Kostečka²,

Pivoňková³

et al. 2011

CAC

Self Cite

View full text Add to dashboard Cite

show abstract

“…In addition to the biotin tags described above, fluorescent dye-hydroxylamine conjugates were also synthesized as covalent probes, which were measured directly for AP site detection [20][21][22]. In a different approach, osmium tetroxide-2,2 -bipyridine was employed as an electrochemical probe of AP sites, which reacts covalently with the unpaired thymine bases in damaged DNA [23]. Furthermore, although a few chemical toxicity sensors based on the detection of oxidized or methylated DNA bases have been developed [24][25][26][27][28][29][30][31][32][33][34][35][36], so far there has been no report on the study of toxicity sensors based on AP site detection.…”

Section: Introductionmentioning

confidence: 99%

A chemical toxicity sensor based on the electrochemiluminescence quantification of apurinic/apyrimidinic sites in double-stranded DNA monolayer

Feng

Liang

Guo

et al. 2017

Sensors and Actuators B: Chemical

View full text Add to dashboard Cite

a b s t r a c tA new chemical toxicity sensor was developed based on the electrochemiluminescence (ECL) quantification of apurinic/apyrimidinic sites (AP sites) in a DNA monolayer with a covalent aldehyde reactive probe (ARP). In the sensor, a uracil-containing DNA duplex was first immobilized on a gold electrode by self-assembly. The duplex was then reacted with uracil-DNA glycosylase (UDG) to convert uracils into AP sites. ARP was employed to tag the AP site with a biotin. After reacting with a ruthenium complex labeled streptavidin, ECL was measured for quantitative analysis. The DNA monolayer was characterized by cyclic voltammetry, electrochemical impedance spectroscopy and chronocoulometry, and its density was measured to be 2.89 × 10 −12 mol/cm 2 . Characterization of the reaction product between ARP and DNA AP sites in solution by nondenaturing polyacrylamide gel electrophoresis and mass spectrometry confirmed successful biotinylation. ECL intensity of the labeled DNA monolayer on the electrode was found to correlate with the number of AP sites, and the detection limit was estimated to be about 1 lesion in 512 DNA bases, which meant that 8.5 fmol AP bases on the electrode were detected. ECL response of the DNA monolayers containing either 8-oxodGuo or methylated bases was very low, indicating that ARP-based AP sites detection method was highly selective. The sensor successfully detected the AP sites in normal DNA induced by methylmethane sulfonate, a carcinogenic chemical. The novel combination of covalent probe and ECL measurement in a sensor configuration therefore provides unique advantages in selectivity and sensitivity, and can be potentially employed in the screening of chemicals for their genetic toxicity.

show abstract

“…The two surface strategy allows to conduct the steps of Os(VIII)-labeling and probe hybridization on magnetic beads, while the detection can be performed on mercury or carbon electrodes [41 -43]. As known from earlier biochemical applications [44], also protective oligonucleotides can be used to label target strands with [OsO 4 [49,50].…”

Section: Introductionmentioning

confidence: 99%

Template‐Free Galvanic Nanostructuring of Gold Electrodes for Sensitive Electrochemical Biosensors

et al. 2009

View full text Add to dashboard Cite

Gold nanostructuring of smooth gold disk-electrodes has been performed by electrodeposition. FE-SEM images indicate a strongly enlarged electrochemically active surface of the modified electrodes. This could be approved by the gold oxide reduction peak in cyclic voltammetry. The electrodes were also applied for sequence-specific DNA detection. For this, complementary single stranded capture probes were immobilized on the electrodes and consecutively hybridized with complementary and noncomplementary [OsO 4 (bipy)]-labeled DNA targets. We found that intermediate dehybridization steps did not decrease the sensitivity for the specific target. Moreover, the surface modification increased the voltammetric signal by an approx. factor of 9. Further, an enhanced linear calibration range was observed. The presented simple nanostructuring process holds great promise for many electrochemical sensor applications.

show abstract

Detection of Abasic Sites in DNA by Electrochemical, Immunoelectrochemical and Acoustic Methods Using OsO₄, 2,2′‐bipyridine as a Probe for Unpaired Thymine Residues

Cited by 17 publications

References 25 publications

Osmium Tetroxide Complexes as Versatile Tools for Structure Probing and Electrochemical Analysis of Biopolymers

Osmium Tetroxide Complexes as Versatile Tools for Structure Probing and Electrochemical Analysis of Biopolymers

A chemical toxicity sensor based on the electrochemiluminescence quantification of apurinic/apyrimidinic sites in double-stranded DNA monolayer

Template‐Free Galvanic Nanostructuring of Gold Electrodes for Sensitive Electrochemical Biosensors

Contact Info

Product

Resources

About

Detection of Abasic Sites in DNA by Electrochemical, Immunoelectrochemical and Acoustic Methods Using OsO4, 2,2′‐bipyridine as a Probe for Unpaired Thymine Residues

Cited by 17 publications

References 25 publications

Osmium Tetroxide Complexes as Versatile Tools for Structure Probing and Electrochemical Analysis of Biopolymers

Osmium Tetroxide Complexes as Versatile Tools for Structure Probing and Electrochemical Analysis of Biopolymers

A chemical toxicity sensor based on the electrochemiluminescence quantification of apurinic/apyrimidinic sites in double-stranded DNA monolayer

Template‐Free Galvanic Nanostructuring of Gold Electrodes for Sensitive Electrochemical Biosensors

Contact Info

Product

Resources

About

Detection of Abasic Sites in DNA by Electrochemical, Immunoelectrochemical and Acoustic Methods Using OsO₄, 2,2′‐bipyridine as a Probe for Unpaired Thymine Residues