title: Redox Sensitivities of Global Cellular Cysteine Residues under Reductive and Oxidative Stress.
abstract: The protein cysteine residue is one of the amino acids most susceptible to oxidative modifications, frequently caused by oxidative stress. Several applications have enabled cysteine-targeted proteomics analysis with simultaneous detection and quantitation. In this study, we employed a quantitative approach using a set of iodoacetyl-based cysteine reactive isobaric tags (iodoTMT) and evaluated the transient cellular oxidation ratio of free and reversibly modified cysteine thiols under DTT and hydrogen peroxide (H2O2) treatments. DTT treatment (1 mM for 5 min) reduced most cysteine thiols, irrespective of their cellular localizations. It also caused some unique oxidative shifts, including for peroxiredoxin 2 (PRDX2), uroporphyrinogen decarboxylase (UROD), and thioredoxin (TXN), proteins reportedly affected by cellular reactive oxygen species production. Modest H2O2 treatment (50 μM for 5 min) did not cause global oxidations but instead had apparently reductive effects. Moreover, with H2O2, significant oxidative shifts were observed only in redox active proteins, like PRDX2, peroxiredoxin 1 (PRDX1), TXN, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Overall, our quantitative data illustrated both H2O2- and reduction-mediated cellular responses, whereby while redox homeostasis is maintained, highly reactive thiols can potentiate the specific, rapid cellular signaling to counteract acute redox stress.
authors.affiliation: Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology , Tokyo 135-0064, Japan.
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Araki
authors.affiliation: Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology , Tokyo 135-0064, Japan.
authors.forename: Hidewo
authors.initials: H
authors.lastname: Kusano
authors.affiliation: Robotic Biology Institute, Inc. , Tokyo 135-0064, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Sasaki
authors.affiliation: Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology , Tokyo 135-0064, Japan.
authors.forename: Riko
authors.initials: R
authors.lastname: Tanaka
authors.affiliation: Robotic Biology Institute, Inc. , Tokyo 135-0064, Japan.
authors.forename: Tomohisa
authors.initials: T
authors.lastname: Hatta
authors.affiliation: Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology , Tokyo 135-0064, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Fukui
authors.affiliation: Molecular Profiling Research Center for Drug Discovery, National Institute of Advanced Industrial Science and Technology , Tokyo 135-0064, Japan.|Robotic Biology Institute, Inc. , Tokyo 135-0064, Japan.
authors.forename: Tohru
authors.initials: T
authors.lastname: Natsume
mesh_terms: D002478:Cells, Cultured; D003545:Cysteine / Q000378:metabolism*; D006706:Homeostasis; D006801:Humans; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D040901:Proteomics; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D013438:Sulfhydryl Compounds; D003545:Cysteine
keywords: cysteine; oxidation; proteomics; quantification; redox