Regular ArticleElectrochemical Determination ofS-Nitrosothiols with a Clark-Type Nitric Oxide Electrode☆
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Aerobic nitric oxide-induced thiol nitrosation in the presence and absence of magnesium cations
2014, Free Radical Biology and MedicineCitation Excerpt :In our standard procedure 2 mM GSH was added to the solution when the NO concentration dropped to ~0.75 µM. After NO had disappeared, 4 mM CuSO4 was added to measure GSNO formation [28]. Where indicated, the concentrations of GSH and MgCl2 were varied between 0.1 and 5 mM and between 0.05 and 20 mM, respectively.
Mechanism and regulation of peroxidase-catalyzed nitric oxide consumption in physiological fluids: Critical protective actions of ascorbate and thiocyanate
2014, Free Radical Biology and MedicineCitation Excerpt :To test whether this increase in NO depletion reflected the formation of S-nitrosoglutathione (GSNO), we examined whether the addition of Cu2+ to decompose S-nitrosothiols could stimulate a characteristic release of NO [25,26]. After MPO-catalyzed NO consumption in the presence of urate and GSH was maximal, the addition of Cu2+ triggered a release of ca. 1 μM NO within 1 min under both aerobic and O2-depleted conditions (Fig. 4D and E), with this being at least equal to the concentration of S-nitrosothiol decomposed [25,26], identifying GSNO as a significant product. Under O2-depleted conditions, this Cu2+-stimulated NO release was sustained and indicated a GSNO yield well in excess of 1 μM (Fig. 4D; under aerobic conditions, NO losses via Cu2+/O2-dependent pathways probably masked this additional NO accumulation [25,26]).
UPLC-MS/MS measurement of S-nitrosoglutathione (GSNO) in human plasma solves the S-nitrosothiol concentration enigma
2013, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :The molar absorptivity coefficient of the S-nitroso group, the most characteristic functional group of RSNO, is quite low (ɛ ≈ 0.8 mM−1 × cm−1 around 334 nm) and does not allow sensitive quantification below about 1 μM by HPLC-UV [4]. Hence, physiological RSNO are commonly measured indirectly, for instance by converting the S-nitroso group to NO or nitrite for detection (for instance Refs. [5–16]). For recent reviews on RSNO analysis see Refs. [17–19]).
S-Nitrosoglutathione
2013, Biochimica et Biophysica Acta - General SubjectsEfficient nitrosation of glutathione by nitric oxide
2013, Free Radical Biology and MedicineCitation Excerpt :Reactions were started by the introduction of DEA/NO (or PROLI/NO, 1 µM) in a total volume of 0.5 ml of 50 mM triethanolamine (TEA) buffer (pH 7.4) and GSH (routinely 1 or 2 mM), 5 mM MgCl2, 1000 U/ml superoxide dismutase (SOD), and 0.1 mM diethylenetriaminepentaacetic acid (DTPA). After complete decay of the NO signal, CuSO4 (4 mM) was added to measure nitrosothiol formation [30]. When indicated, MgCl2 was omitted or replaced by CaCl2, MnCl2, ZnCl2, or NaCl.
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Maines, M. D.
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