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首页> 外文期刊>Electroanalysis >Quantitation of Cu+-catalyzed Decomposition of S-Nitrosoglutathione Using Saville and Electrochemical Detection: a Pronounced Effect of Glutathione and Copper Concentrations
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Quantitation of Cu+-catalyzed Decomposition of S-Nitrosoglutathione Using Saville and Electrochemical Detection: a Pronounced Effect of Glutathione and Copper Concentrations

机译：Saville和电化学检测定量分析Cu +催化S-亚硝基谷胱甘肽的分解：谷胱甘肽和铜浓度的显着影响

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摘要

S-nitrosothiols (RSNOs) are composed of nitric oxide (NO) bound to the sulfhydryl group of amino acids of peptides or proteins. There is a great interest for their quantitation in biological fluids as they have a crucial impact on physiological and pathophysiological events. Most analytical methodologies for quantitation of RSNOs are based on their decomposition followed by the detection of the released NO. In order to obtain the optimal sensitivity for each detection method, the total decomposition of RSNOs is highly desired. The decomposition of RSNOs can be obtained by using catalytically active metal ions, such as Cu+, obtained from CuSO4 in presence of a reducing agent such as glutathione (GSH) that is naturally present in biological environment. In this work, we have re-investigated the decomposition of S-nitrosoglutathione (GSNO) which is the most abundant in vivo low molecular weight RSNO, with a special emphasis on the effect of CuSO4, GSH, and GSNO concentrations and of their ratio. To this aim, GSNO decomposition optimization was performed by both indirect (Griess assay) and direct (real time electrochemical detection of NO at NO-microsensor) quantitation methods. Our results show that the ratio between CuSO4, GSH and GSNO should be adjusted to tune the highest decomposition rate of GSNO and the most efficient electrochemical detection of released NO; also it shows the deleterious effect of very high GSH concentration on the detection of GSNO.
机译：S-亚硝基硫醇（RSNO）由一氧化氮（NO）结合到肽或蛋白质的氨基酸的巯基上组成。由于它们对生理和病理生理事件具有至关重要的影响，因此对它们在生物液体中的定量非常感兴趣。定量RSNO的大多数分析方法都基于其分解，然后检测释放的NO。为了获得每种检测方法的最佳灵敏度，非常需要RSNO的总分解。 RSNO的分解可通过使用催化活性金属离子（例如Cu +）获得，该离子在还原剂（例如谷胱甘肽（GSH））存在下从CuSO4中获得，而该还原剂在生物环境中自然存在。在这项工作中，我们重新研究了S-亚硝基谷胱甘肽（GSNO）的分解，S-亚硝基谷胱甘肽是体内低分子量RSNO含量最高的，特别强调了CuSO4，GSH和GSNO浓度及其比例的影响。为此，通过间接（Griess分析）和直接（在NO微传感器上实时进行NO电化学检测）定量方法进行了GSNO分解优化。我们的结果表明，应调整CuSO4，GSH和GSNO之间的比例，以调节GSNO的最高分解速率和最有效的电化学检测释放的NO。还显示了很高的GSH浓度对GSNO检测的有害作用。

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