ESR detection of intraphagosomal superoxide in polymorphonuclear leukocytes using 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide

Inanami, Osamu; Yamamori, Tohru; Takahashi, Tsuneo; Nagahata, Hajime; Kuwabara, Mikinori

doi:10.1080/10715760100300081

Cited by 18 publications

(4 citation statements)

References 29 publications

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“…A major challenge of radical analysis is the short half-life of superoxide (in the ms to s range) due to its spontaneous dismutation to oxygen and hydrogen peroxide. 10 Chromatography, EPR spin trapping, 11,12 and several dyes 13,14 have been used for the quantification of the radical, along with indirect approaches such as the study of radicalmediated damage of biomolecules. 15,16 Protein electrochemistry has been shown to allow real-time, on-line quantification of radical concentrations.…”

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confidence: 99%

Engineered Superoxide Dismutase Monomers for Superoxide Biosensor Applications

et al. 2005

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Because of its high reaction rate and specificity, the enzyme superoxide dismutase (SOD) offers great potential for the sensitive quantification of superoxide radicals in electrochemical biosensors. In this work, monomeric mutants of human Cu,Zn-SOD were engineered to contain one or two additional cysteine residues, which could be used to bind the protein to gold surfaces, thus making the use of promotor molecules unnecessary. Six mutants were successfully designed, expressed, and purified. All mutants bound directly to unmodified gold surfaces via the sulfur of the cysteine residues and showed a quasi-reversible, direct electron transfer to the electrode. Thermodynamic and kinetic parameters of the electron transfer were characterized and showed only slight variations between the individual mutants. For one of the mutants, the interaction with the superoxide radical was studied in more detail. For both partial reactions of the dismutation, an interaction between protein and radical could be shown. In an amperometric biosensorial approach, the SOD-mutant electrode was successfully applied for the detection of superoxide radicals. In the oxidation region, the electrode surpassed the sensitivity of the commonly used cytochrome c electrodes by approximately 1 order of magnitude while not being limited by interferences, but the electrode did not fully reach the sensitivity of dimeric Cu,Zn-SOD immobilized on MPA-modified gold.

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confidence: 99%

Engineered Superoxide Dismutase Monomers for Superoxide Biosensor Applications

et al. 2005

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“…The stability of their superoxide adducts in vivo or in the presence of biological material remains low and these adducts are easily transformed to diamagnetic compounds. However, most publications dedicated to spin trapping in enzymatic systems, cells or organs make use of these spin traps [10,[17][18][19][20][21][22][23][24] . Recently, cyclic nitrones bearing a triphenylphosphonium group (Mito-DEPMPO and Mito-DIPPMPO, Scheme 1) [25,26] , or conjugated to a -cyclodextrin ring (CDNMPO, CD-DEPMPO, CD-DIPPMPO) [27,28] (Scheme 1) have been synthesized.…”

Section: Introductionmentioning

confidence: 99%

Metabolic stability of superoxide adducts derived from newly developed cyclic nitrone spin traps

Bézière

Hardy

Poulhès

et al. 2014

Free Radical Biology and Medicine

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Reactive oxygen species (ROS) are by-products of aerobic metabolism involved in the onset and evolution of various pathological conditions. Among them, superoxide radical is of special interest as the origin of several damaging species such as H 2 O 2 , hydroxyl radical, or peroxynitrite (ONOO -). Spin trapping coupled with ESR is a method of choice to characterize these species in chemical and biological systems and the metabolic stability of the spin adducts derived from reaction of superoxide and hydroxyl radicals with nitrones is the main limit to the in vivo application of the method. Recently, new cyclic nitrones bearing a triphenylphosphonium or cyclodextrin moiety have been synthesized and their spin adducts demonstrated increased stability in buffer. In the present manuscript, we studied the stability of the superoxide adducts of four new cyclic nitrones in the presence of liver subcellular fractions and biologically relevant reductants using an original set up combining a stoppedflow device and an ESR spectrometer. The kinetics of disappearance of the spin adducts were analyzed using an appropriate simulation program. Our results highlight the interest of new spin trapping agents CD-DEPMPO and CD-DIPPMPO for specific detection of superoxide with high stability of the superoxide adducts in the presence of liver microsomes.3

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“…In this context, the theoretical determination of the DEPMPO-OOH hcc values is a challenging task as the dimensions of this system are prohibitive for high-level wave function-based methods as CCSD or QCISD. Nevertheless, our choice of this system was motivated by the importance of this efficient spin trap in the monitoring of biological processes, ,− like lipid peroxidation inducing DNA or membrane damages. The present study enables us to provide reliable reference hcc values on this realistic system which eventually validate a much cheaper computational approach based on DFT.…”

Section: Introductionmentioning

confidence: 99%

Computation of the Isotropic Hyperfine Coupling Constant: Efficiency and Insights from a New Approach Based on Wave Function Theory

Giner

Tenti

Angeli

et al. 2017

J. Chem. Theory Comput.

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The present paper reports an original computational strategy for the computation of the isotropic hyperfine coupling constants (hcc). The algorithm proposed here is based on an approach recently introduced by some of the authors, namely, the first-order breathing orbital self-consistent field (FOBO-SCF). The approach is an almost parameter-free wave function method capable to accurately treat the spin delocalization together with the spin polarization effects while staying in a restricted formalism and avoiding spin contamination. The efficiency of the method is tested on a series of small radicals, among which four nitroxide radicals and the comparison with high-level ab initio methods show very encouraging results. On the basis of these results, the method is then applied to compute the hcc of a challenging system, namely, the DEPMPO-OOH radical in various conformations. The reference values obtained on such a large system allows us to validate a cheap computational method based on density functional theory (DFT). Another interesting feature of the model applied here is that it allows for the rationalization of the results according to a relatively simple scheme based on a two-step mechanism. More precisely, the results are analyzed in terms of two separated contributions: first the spin delocalization and then the spin polarization.

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ESR detection of intraphagosomal superoxide in polymorphonuclear leukocytes using 5-(diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide

Cited by 18 publications

References 29 publications

Engineered Superoxide Dismutase Monomers for Superoxide Biosensor Applications

Engineered Superoxide Dismutase Monomers for Superoxide Biosensor Applications

Metabolic stability of superoxide adducts derived from newly developed cyclic nitrone spin traps

Computation of the Isotropic Hyperfine Coupling Constant: Efficiency and Insights from a New Approach Based on Wave Function Theory

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