The low-molecular weight biothiols like glutathione (GSH) and cysteine (Cys) play many important roles in various biological processes, the imbalance of biothiols level will lead to many diseases. However, methods that can selectively detect the GSH and Cys was rarely reported because of the similar reactivity and structure. Here, a fluorogenic method was presented to selectively detect the GSH and Cys in vitro using probe ML-FP based on fluorescent protein mimics. Probe ML-FP is a fluorescence turn on probe with lysosome targeting capacity. 2,4-dinitrobenzenesulfonyl serves as fluorescence quench and detection group in probe ML-FP. Differentiating the GSH and Cys was realized benefit from the different reaction time as well as fluorescence response between probe and target species. Low detection limit (4.98 nM for Cys and 4.39 nM for GSH) as well as fast response time was founded in this work. Probe ML-FP possess excellent biocompatibility due to fluorescent protein chromophore and was successfully used for bioimaging in living cells.
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http://dx.doi.org/10.1016/j.talanta.2019.120461 | DOI Listing |
RSC Adv
January 2025
Division of Materials Science, Faculty of Environmental Earth Science, Hokkaido University Japan.
Detecting biothiols like glutathione (GSH), homocysteine (Hcy), and cysteine (Cys) is key to understanding their roles in health and disease. We developed BT-DNBS, a cyanine-based turn-on fluorescent probe with a dinitrobenzenesulfonyl (DNBS) quencher group. Upon biothiol interaction, the quencher is cleaved, restoring fluorescence.
View Article and Find Full Text PDFAnal Chem
January 2025
Department of Chemistry, Indian Institute of Technology Ropar, Rupnagar, Punjab 140001, India.
Thiols, including Cysteine (CYS) and Glutathione (GSH), play pivotal roles in numerous physiological processes as they are integral components of many essential biomolecules and are found abundantly in foods such as additives and antioxidants. Any deviations in thiol concentrations can disrupt normal physiological functions, affecting the body's metabolism and potentially leading to diseases such as Alzheimer's and Parkinson's diseases, etc. Consequently, the imperative need for developing reliable and robust techniques for thiol analysis is crucial for early disease detection and ensuring food safety.
View Article and Find Full Text PDFNat Commun
January 2025
National Institute for Physiological Sciences, National Institutes of Natural Sciences (NINS), Okazaki, Japan.
The presence of redox-active molecules containing catenated sulfur atoms (supersulfides) in living organisms has led to a review of the concepts of redox biology and its translational strategy. Glutathione (GSH) is the body's primary detoxifier and antioxidant, and its oxidized form (GSSG) has been considered as a marker of oxidative status. However, we report that GSSG, but not reduced GSH, prevents ischemic supersulfide catabolism-associated heart failure in male mice by electrophilic modification of dynamin-related protein (Drp1).
View Article and Find Full Text PDFAnal Chem
January 2025
College of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan 410083, China.
Discerning and quantifying the critical biothiols cysteine (Cys), homocysteine (Hcy), and glutathione (GSH) are vital for understanding their synergistic roles in biological systems. In this study, we synthesized a series of phenylethynylcoumarin fluorescent probes with varied structures to investigate the mechanisms underlying biothiol detection. We found that different substituents (-OCH, -H, -CN) at the -position of the phenylacetylene, combined with an aldehyde group at the 3-position of the coumarin, significantly affected the probes' reactivity and produced distinct response patterns toward biothiols.
View Article and Find Full Text PDFFree Radic Biol Med
December 2024
Laboratory of Reproductive Endocrinology, Graduate School of Integrated Sciences for Life, Hiroshima University, 1-4-4, Kagamiyama, Higashihiroshima, Hiroshima, 7398528, Japan. Electronic address:
Sperm cells are highly susceptible to oxidative stress, which decreases their motility and fertility. However, glutathione (GSH) plays a critical role in protecting sperm cells from oxidative damage, a common byproduct of mitochondrial oxidative phosphorylation. On the other hand, GSH biosynthesis in sperm is limited by the availability of cysteine (Cys), which is inherently unstable and found at low concentrations in boar seminal plasma.
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