Abstract
Reactive molecular species (RMS) can damage DNA, lipids, and proteins but as signaling molecules they also affect the regulatory state of the cell. RMS consist of reactive oxygen (ROS), nitrogen (RNS), and carbonyl species (RCS). Besides their potentially destructive nature, RMS are able to modify proteins at the posttranslational level, resulting in regulation of structure, activity, interaction as well as localization. This chapter addresses methods to analyze and quantify posttranslational redox modifications in vitro and ex vivo, such as sulfenic acid generation of cysteine residues and oxidative carbonylation of proteins. In addition, by use of isothermal titration calorimetry, redox-dependent interaction studies of proteins will be described.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Mieyal JJ, Chock PB (2012) Posttranslational modification of cysteine in redox signaling and oxidative stress: focus on s-glutathionylation. Antioxid Redox Signal 16:471–475
Walsh CT, Garneau-Tsodikova S, Gatto GJ (2005) Protein posttranslational modifications: the chemistry of proteome diversifications. Angew Chem Int Ed Engl 44:7342–7372
Seo J-W, Lee K-J (2004) Post-translational modifications and their biological functions: proteomic analysis and aystematic approaches. J Biochem Mol Biol 37:35–44
Mock H-P, Dietz K-J (2016) Redox proteomics for the assessment of redox-related posttranslational regulation in plants. Biochim Biophys Acta 1864:967–973
Woo HA, Kang SW, Kim HK, Yang K-S, Chae HZ, Rhee SG (2003) Reversible oxidation of the active site cysteine of peroxiredoxins to cysteine sulfinic acid. Immunoblot detection with antibodies specific for the hyperoxidized cysteine-containing sequence. J Biol Chem 278:47361–47364
Suzuki YJ, Carini M, Butterfield DA (2010) Protein carbonylation. Antioxid Redox Signal 12:323–325
Lennicke C, Rahn J, Lichtenfels R, Wessjohann LA, Seliger B (2015) Hydrogen peroxide - production, fate and role in redox signaling of tumor cells. Cell Commun Signal 13:39
Bartosz G (2009) Reactive oxygen species: destroyers or messengers? Biochem Pharmacol 77:1303–1315
Gupta V, Carroll KS (2014) Sulfenic acid chemistry, detection and cellular lifetime. Biochim Biophys Acta 1840:847–875
Dalle-Donne I, Rossi R, Giustarini D, Milzani A, Colombo R (2003) Protein carbonyl groups as biomarkers of oxidative stress. Clin Chim Acta 329:23–38
Dalle-Donne I, Scaloni A, Butterfield DA (eds) (2006) Redox proteomics: from protein modifications to cellular dysfunction and diseases, Wiley-interscience series in mass spectrometry. Wiley-Interscience, Hoboken, N.J
Havé M, Leitao L, Bagard M, Castell J-F, Repellin A, Adams W (2015) Protein carbonylation during natural leaf senescence in winter wheat, as probed by fluorescein-5-thiosemicarbazide. Plant Biol (Stuttg) 17:973–979
Levine RL, Williams JA, Stadtman ER, Shacter E (1994) Carbonyl assays for determination of oxidatively modified proteins. Methods Enzymol 233:346–357
Romero-Puertas MC, Palma JM, Gomez M, Del Rio LA, Sandalio LM (2002) Cadmium causes the oxidative modification of proteins in pea plants. Plant Cell Environ 25:677–686
Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG et al (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 186:464–478
Todd MJ, Gomez J (2001) Enzyme kinetics determined using calorimetry: a general assay for enzyme activity? Anal Biochem 296:179–187
Chaires JB (2008) Calorimetry and thermodynamics in drug design. Annu Rev Biophys 37:135–151
Freyer MW, Lewis EA (2008) Isothermal titration calorimetry: experimental design, data dnalysis, and probing macromolecule/ligand binding and kinetic interactions. In: Biophysical tools for biologists, volume one: in vitro techniques, vol 84. Elsevier, Amsterdam; Boston, pp 79–113
Barranco-Medina S, Kakorin S, Lázaro JJ, Dietz K-J (2008) Thermodynamics of the dimer−decamer transition of reduced human and plant 2-Cys peroxiredoxin. Biochemistry 47:7196–7204
Liebthal M, Strüve M, Li X, Hertle Y, Maynard D, Hellweg T et al (2016) Redox-dependent conformational dynamics of decameric 2-cysteine Peroxiredoxin and its interaction with Cyclophilin 20-3. Plant Cell Physiol 57:1415–1425
Matsui K, Sugimoto K, Kakumyan P, Khorobrykh SA, Mano J (2010) Volatile oxylipins and related compounds formed under stress in plants. In: Armstrong D (ed) Lipidomics. Humana Press, Totowa, NJ, pp 17–28
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Treffon, P., Liebthal, M., Telman, W., Dietz, KJ. (2017). Probing Posttranslational Redox Modifications. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 1631. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7136-7_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7136-7_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7134-3
Online ISBN: 978-1-4939-7136-7
eBook Packages: Springer Protocols