title: Anticancer agent 3-bromopyruvic acid forms a conjugate with glutathione.
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.
authors.affiliation: Department of Biochemistry and Cell Biology, Faculty of Biology and Agriculture, University of Rzeszów, Rzeszów, Poland. Electronic address: isadowska@poczta.fm.
authors.forename: Izabela
authors.initials: I
authors.lastname: Sadowska-Bartosz
authors.affiliation: Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Biotechnology and Immunology, Faculty of Biology and Environmental Protection, University of Łódź, Łódź, Poland.
authors.forename: Rafal
authors.initials: R
authors.lastname: Szewczyk
authors.affiliation: Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Germany; Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), Göttingen, Germany.
authors.forename: Lukasz
authors.initials: L
authors.lastname: Jaremko
authors.affiliation: Max-Planck-Institut für Biophysikalische Chemie, Göttingen, Germany.
authors.forename: Mariusz
authors.initials: M
authors.lastname: Jaremko
authors.affiliation: 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.
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Bartosz
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D061986:MCF-7 Cells; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D018384:Oxidative Stress / Q000187:drug effects; D011773:Pyruvates / Q000494:pharmacology*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000970:Antineoplastic Agents; D011773:Pyruvates; C017092:bromopyruvate; D005978:Glutathione
keywords: 3-Bromopyruvate; Erythrocyte; Glutathione; Mass spectrometry