Abstract
Oxidative stress arises when the antioxidant capacity of cells to clean the excess production of reactive oxygen species (ROS) decreases. Several human diseases seem to be related with an increment in the oxidative stress. In this regard, GSH present in the cells works by neutralizing ROS and other xenobiotics through the glutathione S-transferase (GST) enzyme. Thus, the level of expression of GST is an important factor in determining the sensitivity of cells to toxic chemicals or xenobiotic compounds. Therefore, the detection of GST levels is fundamental in the clinical diagnosis of ROS-related diseases. Here, we describe a methodology, based on the voltammetric properties of the ferrocene group (used as electrochemical probe), which can be applied for selective detection of GST levels in human cells. The electrochemical signal measured is associated to the specific interaction of a ferrocenyl-GSH derivate with the G- and H-sites of this enzyme.
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References
Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser F (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 39: 44–84
Quaye IK (2012) Oxidative stress in human health and disease. In: Priti R (ed) Insight and control of infectious diseases in global scenario. InTech, Rijeka, Croatia, pp 97–120
Hayes JD, Flanagan JU, Jowsey IR (2005) Glutathione transferases. Annu Rev Pharmacol Toxicol 45:51–88
Eaton DL, Bammler TK (1999) Concise review of the glutathione S-transferases and their significance to toxicology. Toxicol Sci 49: 156–164
Hayes JD, McLellan LI (1999) Glutathione and glutathione-dependent enzymes represent a co-ordinately regulated defence against oxidative stress. Free Radic Res 31:273–300
Laborde E (2010) Glutathione transferases as mediators of signaling pathways involved in cell proliferation and cell death. Cell Death Differ 17:1373–1380
Fouda MFR, Abd-Elzaher MM, Abdelsamaia RA, Labib AA (2007) On the medicinal chemistry of ferrocene. Appl Organometal Chem 21:613–625
Martos-Maldonado MC, Quesada-Soriano I, García-Maroto F, Vargas-Berenguel A, García-Fuentes L (2012) Ferrocene labelings as inhibitors and dual electrochemical sensors of human glutathione S-transferase p 1-1. Bioorg Med Chem Lett 22:7256–7260
Martos-Maldonado MC, Casas-Solvas JM, Téllez-Sanz R, Mesa-Valle C, Quesada-Soriano I, García-Maroto F, Vargas-Berenguel A, García-Fuentes L (2012) Binding properties of ferrocene-glutathione conjugates as inhibitors and sensors for glutathione S-transferases. Biochimie 94:541–550
Yassin Z, Ortiz-Salmerón E, García-Maroto F, Barón C, García-Fuentes L (2004) Implications of the ligandin binding site on the binding of non-substrate ligands to Schistosoma japonicum-glutathione transferase. Biochim Biophys Acta 1698:227–237
Habig WH, Jakoby WB (1981) Assays for differentiation of glutathione S-transferases. Methods Enzymol 77:398–405
Casas-Solvas JM, Ortiz-Salmerón E, Giménez-Martínez JJ, García-Fuentes L, Capitán-Vallvey LF, Santoyo-González F, Vargas-Berenguel A (2009) Ferrocene-carbohydrate conjugates as electrochemical probes for molecular recognition studies. Chem Eur J 15:710–725
MacDougall D, Crummett WB (1980) Guidelines for data acquisition and data quality evaluation in environmental chemistry. Anal Chem 52:2242–2249
Bard AJ, Faulkner LR (2001) Electrochemical methods. Fundamentals and applications, 2nd edn. Wiley, New York, Sections 6.1, 7.3.4, and 15.6
Wang J (2001) Analytical electrochemistry, 2nd edn. Wiley, New York, Sections 2-1, 3-3.2 and 4-4
Kaifer AE, Gómez-Kaifer M (1999) Supramolecular electrochemistry. Wiley-VCH Verlag GmbH, Weinheim, Sections 1.2, 4.2 and 4.3
Acknowledgments
The authors acknowledge financial support from the Spanish Ministry of Economy and Competitiveness and the EU European Regional Development Fund (Grant CTQ 2010-17848), as well as the Andalusian Regional Government (Consejería de Economía, Innovación y Ciencia, Junta de Andalucía, grants CVI-6028 and FQM-6903).
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Martos-Maldonado, M.C., Casas-Solvas, J.M., Vargas-Berenguel, A., García-Fuentes, L. (2015). Electrochemical Detection of Glutathione S-Transferase: An Important Enzyme in the Cell Protective Mechanism Against Oxidative Stress. In: Armstrong, D. (eds) Advanced Protocols in Oxidative Stress III. Methods in Molecular Biology, vol 1208. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1441-8_10
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DOI: https://doi.org/10.1007/978-1-4939-1441-8_10
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