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Selective targeting of the cysteine proteome by thioredoxin and glutathione redox systems.
Mol Cell Proteomics. 2013 Nov; 12(11):3285-96.MC

Abstract

Thioredoxin (Trx) and GSH are the major thiol antioxidants protecting cells from oxidative stress-induced cytotoxicity. Redox states of Trx and GSH have been used as indicators of oxidative stress. Accumulating studies suggest that Trx and GSH redox systems regulate cell signaling and metabolic pathways differently and independently during diverse stressful conditions. In the current study, we used a mass spectrometry-based redox proteomics approach to test responses of the cysteine (Cys) proteome to selective disruption of the Trx- and GSH-dependent systems. Auranofin (ARF) was used to inhibit Trx reductase without detectable oxidation of the GSH/GSSG couple, and buthionine sulfoximine (BSO) was used to deplete GSH without detectable oxidation of Trx1. Results for 606 Cys-containing peptides (peptidyl Cys) showed that 36% were oxidized more than 1.3-fold by ARF, whereas BSO-induced oxidation of peptidyl Cys was only 10%. Mean fold oxidation of these peptides was also higher by ARF than BSO treatment. Analysis of potential functional pathways showed that ARF oxidized peptides associated with glycolysis, cytoskeleton remodeling, translation and cell adhesion. Of 60 peptidyl Cys oxidized due to depletion of GSH, 41 were also oxidized by ARF and included proteins of translation and cell adhesion but not glycolysis or cytoskeletal remodeling. Studies to test functional correlates showed that pyruvate kinase activity and lactate levels were decreased with ARF but not BSO, confirming the effects on glycolysis-associated proteins are sensitive to oxidation by ARF. These data show that the Trx system regulates a broader range of proteins than the GSH system, support distinct function of Trx and GSH in cellular redox control, and show for the first time in mammalian cells selective targeting peptidyl Cys and biological pathways due to deficient function of the Trx system.

Authors+Show Affiliations

Division of Pulmonary, Allergy and Critical Care Medicine, Department of Medicine.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

23946468

Citation

Go, Young-Mi, et al. "Selective Targeting of the Cysteine Proteome By Thioredoxin and Glutathione Redox Systems." Molecular & Cellular Proteomics : MCP, vol. 12, no. 11, 2013, pp. 3285-96.
Go YM, Roede JR, Walker DI, et al. Selective targeting of the cysteine proteome by thioredoxin and glutathione redox systems. Mol Cell Proteomics. 2013;12(11):3285-96.
Go, Y. M., Roede, J. R., Walker, D. I., Duong, D. M., Seyfried, N. T., Orr, M., Liang, Y., Pennell, K. D., & Jones, D. P. (2013). Selective targeting of the cysteine proteome by thioredoxin and glutathione redox systems. Molecular & Cellular Proteomics : MCP, 12(11), 3285-96. https://doi.org/10.1074/mcp.M113.030437
Go YM, et al. Selective Targeting of the Cysteine Proteome By Thioredoxin and Glutathione Redox Systems. Mol Cell Proteomics. 2013;12(11):3285-96. PubMed PMID: 23946468.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Selective targeting of the cysteine proteome by thioredoxin and glutathione redox systems. AU - Go,Young-Mi, AU - Roede,James R, AU - Walker,Douglas I, AU - Duong,Duc M, AU - Seyfried,Nicholas T, AU - Orr,Michael, AU - Liang,Yongliang, AU - Pennell,Kurt D, AU - Jones,Dean P, Y1 - 2013/08/14/ PY - 2013/8/16/entrez PY - 2013/8/16/pubmed PY - 2014/6/3/medline SP - 3285 EP - 96 JF - Molecular & cellular proteomics : MCP JO - Mol. Cell Proteomics VL - 12 IS - 11 N2 - Thioredoxin (Trx) and GSH are the major thiol antioxidants protecting cells from oxidative stress-induced cytotoxicity. Redox states of Trx and GSH have been used as indicators of oxidative stress. Accumulating studies suggest that Trx and GSH redox systems regulate cell signaling and metabolic pathways differently and independently during diverse stressful conditions. In the current study, we used a mass spectrometry-based redox proteomics approach to test responses of the cysteine (Cys) proteome to selective disruption of the Trx- and GSH-dependent systems. Auranofin (ARF) was used to inhibit Trx reductase without detectable oxidation of the GSH/GSSG couple, and buthionine sulfoximine (BSO) was used to deplete GSH without detectable oxidation of Trx1. Results for 606 Cys-containing peptides (peptidyl Cys) showed that 36% were oxidized more than 1.3-fold by ARF, whereas BSO-induced oxidation of peptidyl Cys was only 10%. Mean fold oxidation of these peptides was also higher by ARF than BSO treatment. Analysis of potential functional pathways showed that ARF oxidized peptides associated with glycolysis, cytoskeleton remodeling, translation and cell adhesion. Of 60 peptidyl Cys oxidized due to depletion of GSH, 41 were also oxidized by ARF and included proteins of translation and cell adhesion but not glycolysis or cytoskeletal remodeling. Studies to test functional correlates showed that pyruvate kinase activity and lactate levels were decreased with ARF but not BSO, confirming the effects on glycolysis-associated proteins are sensitive to oxidation by ARF. These data show that the Trx system regulates a broader range of proteins than the GSH system, support distinct function of Trx and GSH in cellular redox control, and show for the first time in mammalian cells selective targeting peptidyl Cys and biological pathways due to deficient function of the Trx system. SN - 1535-9484 UR - https://www.unboundmedicine.com/medline/citation/23946468/Selective_targeting_of_the_cysteine_proteome_by_thioredoxin_and_glutathione_redox_systems_ L2 - http://www.mcponline.org/cgi/pmidlookup?view=long&pmid=23946468 DB - PRIME DP - Unbound Medicine ER -