Simultaneous detection of small molecule thiols with a simple ¹⁹F NMR platform

Abstract

A 19F NMR platform, capable of discriminating various small molecule thiols, was designed for in-cell thiol differentiation and monitoring, and further detection of the γ-GT activity, demonstrating the wide applications in thiol-related processes.

Small, but powerful and attractive: 19F in biomolecular NMR

“…Using a particular variant, Y125E/Y133E a-synuclein, in which dephosphorylation is slowed down, dephosphorylation and degradation of a-synuclein in real-time could be followed in living cells by fluorine NMR (Zheng et al, 2019). Likewise, real-time monitoring of endogenous thiols in mammalian HeLa cells has been possible, using a tailored fluorinated small molecule reagent, 2,3,5,6-tetrafluroro-4-(phenylsulfonyl)pyridine and derivatives thereof (Chai et al, 2020). For illustration purposes, spectra of 3-F-Tyr-labeled a-synuclein recorded in mammalian cells and cell lysate in my laboratory are provided in Figure 3.…”

Rapid and Selective 19F NMR-Based Sensors for Fingerprint Identification of Ribose

“…Moreover, the lack of naturally occurring fluorinated compounds in the body fluid eliminates background signals and makes the spectrum analysis easier . Consequently, fluorine-labeled molecules combined with 19 F NMR to generate characteristic signals as powerful sensors have been extensively utilized, ranging from organic small molecule discrimination, − biological relevant molecule sensing, − chiral recognition, − drug studies, , and so forth. Furthermore, the combination of fluorinated phenylboronic acid compounds with 19 F NMR has turned out to be a powerful method for sensing of diol-containing analytes. − In particular, a series of fluorinated phenylboronic acid sensor molecules were synthesized by Schiller and co-workers for the identification and discrimination of diols. , Despite remarkable progress, it is still a challenge to identify specific saccharides in a mixture containing multiple potential interferents.…”

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

“…Methods including fluorescence spectroscopy, − spectrophotometry, mass spectrometry, high-performance liquid chromatography, capillary electrophoresis, and Raman spectroscopy have been developed toward detecting and imaging the biothiols in vitro and in cells. Selective determination of individual biothiols generally relies on the intrinsic nucleophilic property of thiols toward the functional group in the probe that has distinct reactivity toward the individual biothiols, of which fluorescence spectrometry was mostly used. − Magnetic resonance including NMR , and EPR spectroscopy is an alternative technique in determining the biothiols. 19 F has similar sensitivity to the proton-based NMR detection, and 19 F NMR has great applications in biological systems. − A few fluorinated probes have been successfully used to distinguish a large variety of thiols in solution and in cells. , Although great efforts have been made to increase the sensitivity in discrimination of biothiols, simultaneous identification and quantification of the four biothiols (Cys, GSH, Hcy, and H 2 S) close to the physiological conditions are still a challenge.…”

“…Selective determination of individual biothiols generally relies on the intrinsic nucleophilic property of thiols toward the functional group in the probe that has distinct reactivity toward the individual biothiols, of which fluorescence spectrometry was mostly used. 21−29 Magnetic resonance including NMR 30,31 and EPR spectroscopy 32 is an alternative technique in determining the biothiols. 19 F has similar sensitivity to the proton-based NMR detection, 33 and F NMR has great applications in biological systems.…”