Abstract
S-Nitrosation is a key posttranslational modification in regulating proteins in both normal physiology and diverse human diseases. To identify novel therapies for human diseases linked to oxidative and nitrosative stress, understanding how cells control S-nitrosation specificity could be critical. Among the enzymes known to control S-nitrosation of proteins, thioredoxin 1 (Trx1), a conserved disulfide reductase, transnitrosates and denitrosates distinct sets of target proteins. To recognize the function of Trx1 in both normal and dysfunctional cells, S-nitrosation targets of Trx1 in different cells need to be identified. However, S-nitrosation is usually too labile to be detected directly by mass spectrometry (MS). Here we present two optimized MS techniques to identify S-nitrosated Trx1 and its transnitrosation targets, using both direct and indirect MS methods.
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References
Foster MW, Hess DT, Stamler JS (2009) Protein S-nitrosylation in health and disease: a current perspective. Trends Mol Med 15(9):391–404
Mannick JB (2007) Regulation of apoptosis by protein S-nitrosylation. Amino Acids 32(4):523–526
Benhar M, Forrester MT, Hess DT, Stamler JS (2008) Regulated protein denitrosylation by cytosolic and mitochondrial thioredoxins. Science 320(5879):1050–1054
Gao C, Guo H, Wei J, Mi Z, Wai PY, Kuo PC (2005) Identification of S-nitrosylated proteins in endotoxin-stimulated RAW264.7 murine macrophages. Nitric Oxide 12(2):121–126
Wu C, Liu T, Chen W, Oka S, Fu C, Jain MR, Parrott AM, Baykal AT, Sadoshima J, Li H (2010) Redox regulatory mechanism of transnitrosylation by thioredoxin. Mol Cell Proteomics 9(10):2262–2275
Haendeler J, Hoffmann J, Tischler V, Berk BC, Zeiher AM, Dimmeler S (2002) Redox regulatory and anti-apoptotic functions of thioredoxin depend on S-nitrosylation at cysteine 69. Nat Cell Biol 4(10):743–749
Haendeler J, Weiland U, Zeiher AM, Dimmeler S (1997) Effects of redox-related congeners of NO on apoptosis and caspase-3 activity. Nitric Oxide 1(4):282–293
Hashemy SI, Holmgren A (2008) Regulation of the catalytic activity and structure of human thioredoxin 1 via oxidation and S-nitrosylation of cysteine residues. J Biol Chem 283(32):21890–21898
Wu C, Parrott AM, Fu C, Liu T, Marino SM, Gladyshev VN, Jain MR, Baykal AT, Li Q, Oka S, Sadoshima J, Beuve A, Simmons WJ, Li H (2011) Thioredoxin 1-mediated post-translational modifications: reduction, transnitrosylation, denitrosylation, and related proteomics methodologies. Antioxid Redox Signal 15(9):2565–2604
Greco TM, Hodara R, Parastatidis I, Heijnen HF, Dennehy MK, Liebler DC, Ischiropoulos H (2006) Identification of S-nitrosylation motifs by site-specific mapping of the S-nitrosocysteine proteome in human vascular smooth muscle cells. Proc Natl Acad Sci U S A 103(19):7420–7425
Hao G, Derakhshan B, Shi L, Campagne F, Gross SS (2006) SNOSID, a proteomic method for identification of cysteine S-nitrosylation sites in complex protein mixtures. Proc Natl Acad Sci U S A 103(4):1012–1017
Jaffrey SR, Erdjument-Bromage H, Ferris CD, Tempst P, Snyder SH (2001) Protein S-nitrosylation: a physiological signal for neuronal nitric oxide. Nat Cell Biol 3(2):193–197
Paige JS, Xu G, Stancevic B, Jaffrey SR (2008) Nitrosothiol reactivity profiling identifies S-nitrosylated proteins with unexpected stability. Chem Biol 15(12):1307–1316
Torta F, Usuelli V, Malgaroli A, Bachi A (2008) Proteomic analysis of protein S-nitrosylation. Proteomics 8(21):4484–4494
Gow AJ, Davis CW, Munson D, Ischiropoulos H (2004) Immunohistochemical detection of S-nitrosylated proteins. Methods Mol Biol 279:167–172
Jaffrey SR, Snyder SH (2001) The biotin switch method for the detection of S-nitrosylated proteins. Sci STKE 2001(86):PL1
Huang B, Chen C (2006) An ascorbate-dependent artifact that interferes with the interpretation of the biotin switch assay. Free Radic Biol Med 41(4):562–567
Knipp M, Braun O, Gehrig PM, Sack R, Vasák M (2003) Zn(II)-free dimethylargininase-1 (DDAH-1) is inhibited upon specific Cys-S-nitrosylation. J Biol Chem 278(5):3410–3416
Wang Y, Liu T, Wu C, Li H (2008) A strategy for direct identification of protein S-nitrosylation sites by quadrupole time-of-flight mass spectrometry. J Am Soc Mass Spectrom 19(9):1353–1360
Lee SJ, Lee JR, Kim YH, Park YS, Park SI, Park HS, Kim KP (2007) Investigation of tyrosine nitration and nitrosylation of angiotensin II and bovine serum albumin with electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom 21(17):2797–2804
Wu C, Parrott AM, Liu T, Beuve A, Li H (2013) Functional proteomics approaches for the identification of transnitrosylase and denitrosylase targets. Methods 62(2):151–160
Wu C, Parrott AM, Liu T, Jain MR, Yang Y, Sadoshima J, Li H (2011) Distinction of thioredoxin transnitrosylation and denitrosylation target proteins by the ICAT quantitative approach. J Proteome 74(11):2498–2509
Acknowledgments
This chapter is produced with a grant support from the NIH-National Institute of General Medical Sciences (R01GM112415, to A.B. and H.L.), and the Orbitrap MS described here was purchased with a grant from the NIH-National Institute of Neurological Disorders and Stroke (P30NS046593).
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Wu, C. et al. (2018). Biotin Switch Processing and Mass Spectrometry Analysis of S-Nitrosated Thioredoxin and Its Transnitrosation Targets. In: Mengel, A., Lindermayr, C. (eds) Nitric Oxide. Methods in Molecular Biology, vol 1747. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7695-9_20
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DOI: https://doi.org/10.1007/978-1-4939-7695-9_20
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