Izabela Sadowska-Bartosz1, Rafal Szewczyk2, Lukasz Jaremko3, Mariusz Jaremko4, Grzegorz Bartosz5. 1. Department of Biochemistry and Cell Biology, Faculty of Biology and Agriculture, University of Rzeszów, Rzeszów, Poland. Electronic address: isadowska@poczta.fm. 2. Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland. 3. Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Germany; Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Göttingen, Germany. 4. Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Germany. 5. Department of Biochemistry and Cell Biology, Faculty of Biology and Agriculture, University of Rzeszów, Rzeszów, Poland; Department of Molecular Biophysics, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland.
Abstract
BACKGROUND: 3-Bromopyruvic acid (3-BP), a glycolytic inhibitor and a promising anticancer compound, induces oxidative stress and depletes cells of glutathione (GSH). The causes of GSH loss remain unclear. The aim of this study was to ascertain whether 3-BP forms a conjugate with glutathione. METHODS: GSH was incubated with various amounts of 3-BP and the extent of reaction was titrated with (1)H NMR and (1)H-(1)H NMR. The reaction outcome was identified by MS/MS. Intracellular formation of the conjugate was assessed in cells treated with 3-BP and 3-BP((13)C) and analyzed using the targeted LC-MS/MS method in negative ionization MRM mode. RESULTS: 3-BP was found to react with GSH in a 1:1 ratio forming an S-conjugate. The same conjugate was formed intracellularly in erythrocytes and MCF-7 cells. CONCLUSIONS: 3-BP reacts with GSH in the absence of cells and intracellularly. This reaction appears to be the main cause of GSH loss in 3-BP treated cells.
BACKGROUND:3-Bromopyruvic acid (3-BP), a glycolytic inhibitor and a promising anticancer compound, induces oxidative stress and depletes cells of glutathione (GSH). The causes of GSH loss remain unclear. The aim of this study was to ascertain whether 3-BP forms a conjugate with glutathione. METHODS:GSH was incubated with various amounts of 3-BP and the extent of reaction was titrated with (1)H NMR and (1)H-(1)H NMR. The reaction outcome was identified by MS/MS. Intracellular formation of the conjugate was assessed in cells treated with 3-BP and 3-BP((13)C) and analyzed using the targeted LC-MS/MS method in negative ionization MRM mode. RESULTS:3-BP was found to react with GSH in a 1:1 ratio forming an S-conjugate. The same conjugate was formed intracellularly in erythrocytes and MCF-7 cells. CONCLUSIONS:3-BP reacts with GSH in the absence of cells and intracellularly. This reaction appears to be the main cause of GSH loss in 3-BP treated cells.
Authors: Minjong Lee; Ara Jo; Seulki Lee; Jong Bin Kim; Young Chang; Joon Yeul Nam; Hyeki Cho; Young Youn Cho; Eun Ju Cho; Jeong-Hoon Lee; Su Jong Yu; Jung-Hwan Yoon; Yoon Jun Kim Journal: PLoS One Date: 2017-03-31 Impact factor: 3.240
Authors: Irini Skaripa-Koukelli; David Hauton; John Walsby-Tickle; Eloïse Thomas; Joshua Owen; Abirami Lakshminarayanan; Sarah Able; James McCullagh; Robert C Carlisle; Katherine A Vallis Journal: Cancer Metab Date: 2021-10-14