Selective Disruption of Mitochondrial Thiol Redox State in Cells and In Vivo
| Autor: | Michael P. Murphy, Andrew M. James, Thomas P. Bright, Andrew R. Hall, Angela Logan, Hiran A. Prag, Elizabeth C. Hinchy, Lee M. Booty, Cristiane Benincá, Julien Prudent, Tracy A. Prime, Menna R. Clatworthy, John F. Mulvey, Justyna M. Gawel, Thomas Krieg, Filip Cvetko, Stuart T. Caldwell, John R. Ferdinand, Richard C. Hartley, Sabine Arndt |
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| Přispěvatelé: | Clatworthy, Menna [0000-0002-3340-9828], Ferdinand, John [0000-0003-0936-0128], Prag, Hiran [0000-0002-4753-8567], Prudent, Julien [0000-0003-3821-6088], Krieg, Thomas [0000-0002-5192-580X], Murphy, Mike [0000-0003-1115-9618], Apollo - University of Cambridge Repository |
| Rok vydání: | 2019 |
| Předmět: |
Clinical Biochemistry
Cell Mitochondrion Biology 01 natural sciences Biochemistry Article Mice chemistry.chemical_compound Thioredoxins Tandem Mass Spectrometry Drug Discovery Organelle Dinitrochlorobenzene medicine Animals Humans Sulfhydryl Compounds glutathione redox signaling Molecular Biology Chromatography High Pressure Liquid Glutathione Transferase Membrane Potential Mitochondrial Pharmacology chemistry.chemical_classification Reactive oxygen species 010405 organic chemistry mitochondria targeting Hep G2 Cells thioredoxin Glutathione Recombinant Proteins 0104 chemical sciences Cell biology mitochondria Cytosol medicine.anatomical_structure Liver chemistry Molecular Medicine thiol redox state Thioredoxin Reactive Oxygen Species Peroxiredoxin Oxidation-Reduction |
| Zdroj: | Cell Chemical Biology |
| ISSN: | 2451-9456 |
| DOI: | 10.1016/j.chembiol.2018.12.002 |
| Popis: | Summary Mitochondrial glutathione (GSH) and thioredoxin (Trx) systems function independently of the rest of the cell. While maintenance of mitochondrial thiol redox state is thought vital for cell survival, this was not testable due to the difficulty of manipulating the organelle's thiol systems independently of those in other cell compartments. To overcome this constraint we modified the glutathione S-transferase substrate and Trx reductase (TrxR) inhibitor, 1-chloro-2,4-dinitrobenzene (CDNB) by conjugation to the mitochondria-targeting triphenylphosphonium cation. The result, MitoCDNB, is taken up by mitochondria where it selectively depletes the mitochondrial GSH pool, catalyzed by glutathione S-transferases, and directly inhibits mitochondrial TrxR2 and peroxiredoxin 3, a peroxidase. Importantly, MitoCDNB inactivates mitochondrial thiol redox homeostasis in isolated cells and in vivo, without affecting that of the cytosol. Consequently, MitoCDNB enables assessment of the biomedical importance of mitochondrial thiol homeostasis in reactive oxygen species production, organelle dynamics, redox signaling, and cell death in cells and in vivo. Graphical Abstract Highlights • MitoCDNB is a mitochondria-targeted molecule that disrupts thiol redox state • The CDNB moiety depletes glutathione and inhibits key thiol redox enzymes • MitoCDNB selectively disrupts mitochondrial thiol redox state in cells and in vivo • MitoCDNB extends methods available to investigate mitochondrial thiol redox state It has been difficult to selectively disrupt mitochondrial redox state, independently of that in the rest of the cell. Here, Booty et al. introduce MitoCDNB to deplete mitochondrial glutathione and selectively inhibit key enzymatic components of mitochondrial thiol redox status in cells and in vivo. |
| Databáze: | OpenAIRE |
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