Simultaneous Quantification of Biothiols and Deciphering Diverse GSH Stability in Different Live Cells by 19F-Tag,Analytical Chemistry

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Simultaneous Quantification of Biothiols and Deciphering Diverse GSH Stability in Different Live Cells by 19F-Tag
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-12-27 , DOI: 10.1021/acs.analchem.1c03673
Chao-Yu Cui ₁ , Bin Li ₁ , Dan Cheng ₁ , Xia-Yan Li ₁ , Jia-Liang Chen ₁ , Ya-Ting Chen ₁ , Xun-Cheng Su ₁

Affiliation

GSH, Cys, Hcy, and H₂S are important biothiols and play important roles in the living systems. Quantitative and simultaneous determination of these biothiols under physiological conditions is still a challenge. Herein, we developed an effective ¹⁹F-reactive tag that readily interacts with these four biothiols for the generation of stable thioether products that have distinguishable ¹⁹F-chemical shifts. These thioester compounds encode the characteristic fingerprint profiles of each biothiols, allowing one to simultaneously quantify and determine these biothiols by 1D ¹⁹F NMR spectroscopy. The intra-/extracellular GSH in live cells was assessed by the established strategy, and remarkable variations in the GSH stability were determined between the normal mammalian cells and cancer cells. It is notable that GSH hydrolyzes efficiently in the out-membrane of the cancer cells and the lysates. In contrast, GSH remains stable in the tested normal cells.

中文翻译：

通过 19F-Tag 同时定量生物硫醇和破译不同活细胞中不同的 GSH 稳定性

GSH、Cys、Hcy和H ₂ S是重要的生物硫醇，在生命系统中发挥着重要作用。在生理条件下定量和同时测定这些生物硫醇仍然是一个挑战。在这里，我们开发了一种有效的¹⁹ F 反应性标签，该标签很容易与这四种生物硫醇相互作用，以生成具有可区分的¹⁹ F 化学位移的稳定硫醚产品。这些硫酯化合物对每种生物硫醇的特征指纹图谱进行编码，从而可以通过 1D ^{19同时量化和确定这些生物硫醇}F核磁共振波谱。通过既定策略评估活细胞中的细胞内/细胞外 GSH，并确定正常哺乳动物细胞和癌细胞之间 GSH 稳定性的显着变化。值得注意的是，GSH 在癌细胞和裂解物的膜外有效水解。相反，GSH 在测试的正常细胞中保持稳定。

更新日期：2022-01-18

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