title: Detection of Cysteine Redox States in Mitochondrial Proteins in Intact Mammalian Cells.
abstract: Import, folding, and activity regulation of mitochondrial proteins are important for mitochondrial function. Cysteine residues play crucial roles in these processes as their thiol groups can undergo (reversible) oxidation reactions. For example, during import of many intermembrane space (IMS) proteins, cysteine oxidation drives protein folding and translocation over the outer membrane. Mature mitochondrial proteins can undergo changes in the redox state of specific cysteine residues, for example, as part of their enzymatic reaction cycle or as adaptations to changes of the local redox environment which might influence their activity. Here we describe methods to study changes in cysteine residue redox states in intact cells. These approaches allow to monitor oxidation-driven protein import as well as changes of cysteine redox states in mature proteins during oxidative stress or during the reaction cycle of thiol-dependent enzymes like oxidoreductases.
authors.affiliation: Institute for Biochemistry, University of Cologne, Zuelpicher Str 47a, 50674, Cologne, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Habich
authors.affiliation: Institute for Biochemistry, University of Cologne, Zuelpicher Str 47a, 50674, Cologne, Germany. jan.riemer@uni-koeln.de.
authors.forename: Jan
authors.initials: J
authors.lastname: Riemer
mesh_terms: D002458:Cell Fractionation / Q000379:methods; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism*; D004220:Disulfides; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D047468:Immunoprecipitation; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D017510:Protein Folding; D021381:Protein Transport
chemical_list: D004220:Disulfides; D024101:Mitochondrial Proteins; D003545:Cysteine
keywords: Cysteine; Intermembrane space (IMS); Mia40; Oxidative folding; Redox state; Thiol modification