Abstract
Redox signaling in the cell, which is essential for cell physiology, involves proteins with free sulfhydryl groups (−SH). Among them, the thioredoxin system plays the most significant role. Many conditions associated with cell malignancies feature the higher expression of thioredoxin, making it an attractive target for new therapeutic drug development. Here we present a simple in vitro model of testing the interaction between thioredoxin and the putative drug. This method is relatively inexpensive and gives the Investigator a first screen of the drug properties, which can be essential for further experimental approaches.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kozakowska M, Pietraszek-Gremplewicz K, Jozkowicz A et al (2015) The role of oxidative stress in skeletal muscle injury and regeneration: focus on antioxidant enzymes. J Muscle Res Cell Motil 36:377–393
Kirkinezos I, Moraes CT (2001) Reactive oxygen species and mitochondrial diseases. Cell Dev Biol 12:449–457
Majima HJ, Indo HP, Nakanishi I et al (2016) Chasing great paths of Helmut Sies “oxidative stress”. Arch Biochem Biophys 595:54–60
Yin F, Sancheti H, Patil I et al (2016) Energy metabolism and inflammation in brain aging and Alzheimer’s disease. Free Radic Biol Med 16:30216–30217
Wright E Jr, Scism-Bacon JL, Glass LC (2006) Oxidative stress in type 2 diabetes: role of fasting and postprandial glycaemia. Int J Clin Pract 60:308–314
De Marchi E, Baldassari F, Bononi A et al (2013) Oxidative stress in cardiovascular disease and obesity: role of p66 Shc and protein C. Oxid Med Cell Longer
Klaunig JE, Kamendulis LM, Hovecar BA (2010) Oxidative stress and oxidative damage in carcinogenesis. Toxicol Pathol 38:96–109
Li W, Kong AN (2008) Molecular mechanism of Nrf2-mediated antioxidant response. Mol Carcinog 47:91–104
Schumacker PT (2006) Reactive oxygen species in cancer cells: live by the sword, die by the sword. Cancer Cell 10:175–176
Forman HJ, Fukuto JM, Torres M (2000) Redox signaling: thiol chemistry defines which reactive oxygen and nitrogen species can act as a second messenger. Am J Physiol Cell Physiol 287:C246–C256
Adimora NJ, Jones DP, Kemp ML (2010) A model of redox kinetics implicates the thiol proteome in cellular hydrogen peroxide response. Antioxid Redox Signal 13:731–743
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 283:21890–21898
Eklund H, Gleason FK, Holmgren A (1991) Structural and functional relations among thioredoxins of different species. Proteins 11(13):13–28
Chivers PT, Prehoda KE, Volkman BF et al (1997) Microscopic pK(a) values of Escherichia coli thioredoxin. Biochemistry 36(15):14985–11499
Messens J, Van Molle I, Vanhaesebrouck P et al (2004) How thioredoxin can reduce a buried disulphide bonds. J Mol Biol 339(16):527–537
Das KC, Das CK (2000) Thioredoxin, a singlet oxygen quencher and hydroxyl radical scavenger: redox independent functions. Biochem Biophys Res Commun 277(17):443–447
Winterbourn CC, Hampton MB (2008) Thiol chemistry and specificity in redox signaling. Free Radic Biol Med 45(18):549–561
Depuydt M, Leonard SE, Vertommen D et al (2009) A periplasmic reducing systems protects single cysteine residues from oxidation. Science 20(19):1109–1111
Lu J, Holmgren A (2014) Thiredoxin superfamily in oxidative protein folding. Antioxid Redox Signal 21(20):457–470
Laurent TC, Moore EC, Reichard P (1964) Enzymatic synthesis of deoxyribonucleotides. IV. Isolation and characterization of thioredoxin, the hydrogen donor from Escherichia coli B. J Biol Chem 239(21):3436–3444
Matthews JR, Wakasugi N, Virelizier JL (1992) Thioredoxin regulates the DNA binding activity of NF-κB by reduction of disulfide bond involving cysteine 62. Nucleic Acid Res 20(22):3821–3830
Saitoh M, Nishitoh H, Fujii M (1998) Mammalian thioredoxin is a direct inhibitor of apoptosis signal-regulating kinase (ASK). EMBO J 17(23):2596–2606
Zhang D, Ahao N, Ma B et al (2016) Procaspase-9 induces its cleavage by transnitrosilating XIAP via thioredoxin system during cerebral ischemia-reperfusion in rats. Sci Rep 6
Matsui M, Oshima M, Oshima H et al (1996) Early embryonic lethality caused by targeted disruption of the mouse thioredoxin gene. Dev Biol 178(25):179–185
Nonn L, Williams RR, Erickson RP et al (2003) The absence of thioredoxin 2 causes massive apoptosis, exencephaly, and early embryonic development in homozygous mice. Mol Cell Biol 23(26):916–922
Fontaine SD, Reid R, Robinson L et al (2015) Long-term stabilization of maleimide-thiol conjugates. Bioconjug Chem 26(27):145–152
Skalska J, Brookes PS, Nadtochiy S et al (2009) Modulation of cell surface protein free thiols: a potential novel mechanism of action of the sesquiterpene lactone parthenolide. Plos One 4(28):8115–8127
Shafer D, Inaman JK, Lees A (2000) Reaction of Tris(2-carboxyethyl)phosphine (TCEP) with maleimide and α-halocyl groups: anomalous elution of TCEP by gel filtration. Anal Biochem 282(29):161–164
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Skalska, J. (2019). Thioredoxin-1 PEGylation as an In Vitro Method for Drug Target Identification. In: Hancock, J., Conway, M. (eds) Redox-Mediated Signal Transduction. Methods in Molecular Biology, vol 1990. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9463-2_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9463-2_12
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9461-8
Online ISBN: 978-1-4939-9463-2
eBook Packages: Springer Protocols