Profiling thiol metabolites and quantification of cellular glutathione using FT-ICR-MS spectrometry.
Profiling thiol metabolites and quantification of cellular glutathione using FT-ICR-MS spectrometry.
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DOI：
10.1007/s00216-014-7810-z
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发表时间：
2014-07
影响因子：
4.3
通讯作者：
Nantz, Michael H.
中科院分区：
文献类型：
--
作者：
Gori, Sadakatali S.;Lorkiewicz, Pawel;Ehringer, Daniel S.;Belshoff, Alex C.;Higashi, Richard M.;Fan, Teresa W. -M.;Nantz, Michael H.
本文报道了季铵基α-碘乙酰胺QDE及其同位素体 *QDE的制备及其作为细胞硫醇化学选择性衍生试剂的应用。将试剂直接加入到活细胞中，然后加合物萃取到正丁醇中并通过FT-ICR-MS进行分析，提供了一个匹配同位素峰的注册表，从中推导出硫醇代谢物的分子式。在细胞裂解和加合物形成过程中酸化至pH 4进一步提高了硫醇衍生化的化学选择性。使用这种方法检查A549人肺腺癌细胞，发现半胱氨酸，半胱氨酰甘氨酸，谷胱甘肽和高半胱氨酸作为主要的巯基代谢物以及亚磺酸亚牛磺酸。该方法也易于用于定量硫醇代谢物，如本文通过分别在34.4±11.5和10.1± 4.0 nmol/mg蛋白质浓度下定量A549细胞中的谷胱甘肽和谷胱甘肽二硫化物所证明的。
We describe preparation and use of the quaternary ammonium-based α-iodoacetamide QDE and its isotopologue *QDE as reagents for chemoselective derivatization of cellular thiols. Direct addition of the reagents to live cells followed by adduct extraction into n-butanol and analysis by FT-ICR-MS provided a registry of matched isotope peaks from which molecular formulae of thiol metabolites were derived. Acidification to pH 4 during cell lysis and adduct formation further improves the chemoselectivity for thiol derivatization. Examination of A549 human lung adenocarcinoma cells using this approach revealed cysteine, cysteinylglycine, glutathione, and homocysteine as principal thiol metabolites as well as the sulfinic acid hypotaurine. The method is also readily applied to quantify the thiol metabolites, as demonstrated here by the quantification of both glutathione and glutathione disulfide in A549 cells at concentrations of 34.4±11.5 and 10.1± 4.0 nmol/mg protein, respectively.
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影响因子：
5.3
作者：
Bošković M;Vovk T;Kores Plesničar B;Grabnar I
通讯作者：
Grabnar I
使用色烯分子对硫醇进行比色检测
影响因子：
5.2
作者：
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影响因子：
6.2
作者：
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影响因子：
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作者：
Hill, Bradford G.;Reily, Colin;Oh, Joo-Yeun;Johnson, Michelle S.;Landar, Aimee
通讯作者：
Landar, Aimee