DNA sensing by electrocatalysis with hemoglobin

Pheeney, Catrina G.; Guerra, Luis F.; Barton, Jacqueline K.

doi:10.1073/pnas.1201551109

Cited by 35 publications

(31 citation statements)

References 43 publications

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“…Also, hemoglobin exhibits the same feature and has been shown to couple with Nile blue. 58 Detection platforms based on these redox-active proteins are appealing due to their inherent catalytic activity. In addition, the application of a protein as an electron sink is highly attractive due to the ability of the protein shell to shield the redox core from the electrode surface.…”

Section: Enhancement Of Sensmentioning

confidence: 99%

Protein Electrocatalysis for Direct Sensing of Circulating MicroRNAs

2014

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MicroRNAs (miRNAs) are potentially useful biomarkers for diagnosis, classification, and prognosis of many diseases, including cancer. Herein, we developed a protein-facilitated electrocatalytic quadroprobe sensor (Sens(PEQ)) for detection of miRNA signature of chronic lymphocytic leukemia (CLL) in human serum. The developed signal-ON sensor provides a compatible combination of two DNA adaptor strands modified with four methylene blue molecules and electrocatalysis using glucose oxidase in order to enhance the overall signal gain. This enhanced sensitivity provided the response necessary to detect the low-abundant serum miRNAs without preamplification. The developed Sens(PEQ) is exquisitely sensitive to subtle π-stack perturbations and capable of distinguishing single base mismatches in the target miRNA. Furthermore, the developed sensor was employed for profiling of three endogenous miRNAs characteristic to CLL, including hsa-miR-16-5p, hsa-miR-21-5p, and hsa-miR-150-5p in normal healthy serum, chronic lymphocytic leukemia Rai stage 1 (CLL-1), and stage 3 (CLL-3) sera, using a non-human cel-miR-39-3p as an internal standard. The sensor results were verified by conventional SYBR green-based quantitative reverse-transcription polymerase chain reaction (RT-qPCR) analysis.

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Section: Enhancement Of Sensmentioning

confidence: 99%

Protein Electrocatalysis for Direct Sensing of Circulating MicroRNAs

2014

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“…Because of the iron center shielding afforded by hemoglobin’s native protein conformation, passivation of the DNA-modified electrode is less of a stringent necessity when this protein is used as an electron sink (Figure 7) [36]. The combination of a covalent MB mediator and hemoglobin electron sink enabled the ready detection of restriction-enzyme activity at a low-density DNA film.…”

Section: Electrocatalysis With Dna Ct For Dna Sensingmentioning

confidence: 99%

Electrocatalysis in DNA sensors

2014

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Electrocatalysis is often thought of solely in the inorganic realm, most often applied to energy conversion in fuel cells. However, the ever-growing field of bioelectrocatalysis has made great strides in advancing technology for both biofuel cells as well as biological detection platforms. Within the context of bioelectrocatalytic detection systems, DNA-based platforms are especially prevalent. One subset of these platforms, the one we have developed, takes advantage of the inherent charge transport properties of DNA. Electrocatalysis coupled with DNA-mediated charge transport has enabled specific and sensitive detection of lesions, mismatches and DNA-binding proteins. Even greater signal amplification from these platforms is now being achieved through the incorporation of a secondary electrode to the platform both for patterning DNA arrays and for detection. Here, we describe the evolution of this new DNA sensor technology.

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“…Multiplexed DNA-modified electrodes have been developed to sense an extensive range of targets including small molecule (4, 5, 11), DNA (6, 10, 11, 12), RNA (8, 9, 11), and proteins (3, 7, 11–13). These devices all strive to achieve the same goals of enhanced sensitivity, faster detection times, and tolerance to cell lysates (5, 9, 12). Despite variety in the design of these multiplexed DNA-modified electrodes, they all possess the same intrinsic benefits of statistical comparisons and parallel experimentation; these advantages have proven to be essential for the electrochemical characterization of complex systems.…”

Section: Introductionmentioning

confidence: 99%

Multiplexed Electrochemistry of DNA-Bound Metalloproteins

et al. 2013

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Here we describe a multiplexed electrochemical characterization of DNA-bound proteins containing [4Fe-4S] clusters. DNA-modified electrodes have become an essential tool for the characterization of the redox chemistry of DNA repair proteins containing redox cofactors, and multiplexing offers a means to probe different complex samples and substrates in parallel to elucidate this chemistry. Multiplexed analysis of EndonucleaseIII (EndoIII), a DNA repair protein containing a [4Fe-4S] cluster known to be accessible via DNA-mediated charge transport, shows subtle differences in the electrochemical behavior as a function of DNA morphology. The peak splitting, signal broadness, sensitivity to π-stack perturbations, and kinetics were all characterized for the DNA-bound reduction of EndoIII on both closely and loosely packed DNA films. DNA-bound EndoIII is seen to have two different electron transfer pathways for reduction, either through the DNA base stack or through direct surface reduction; closely packed DNA films, where the protein has limited surface accessibility, produce electrochemical signals reflecting electron transfer that is DNA-mediated. Multiplexing furthermore permits the comparison of the electrochemistry of EndoIII mutants, including a new family of mutations altering the electrostatics surrounding the [4Fe-4S] cluster. While little change in the midpoint potential was found for this family of mutants, significant variations in the efficiency of DNA-mediated electron transfer were apparent. Based on the stability of these proteins, examined by circular dichroism, we propose that the electron transfer pathway can be perturbed not only by the removal of aromatic residues but also through changes in solvation near the cluster.

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DNA sensing by electrocatalysis with hemoglobin

Cited by 35 publications

References 43 publications

Protein Electrocatalysis for Direct Sensing of Circulating MicroRNAs

Protein Electrocatalysis for Direct Sensing of Circulating MicroRNAs

Electrocatalysis in DNA sensors

Multiplexed Electrochemistry of DNA-Bound Metalloproteins

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