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The Influence of Some Polyphenols on Human Erythrocytes Glutathione S-Transferase Activity

Authors: Mustafa Erat


Glutathione S-transferase was purified from human erythrocytes and effects of some polyphenols were investigated on the enzyme activity. The purification procedure was performed on Glutathione-Agarose affinity chromatography after preparation of erythrocytes hemolysate with a yield of 81%. The purified enzyme showed a single band on the SDS-PAGE. The effects of some poliphenolic compounds such as catechin, dopa, dopamine, progallol and catechol were examined on the in vitro GST activity. Catechin was determined to be inhibitor for the enzyme, but others were not effective on the enzyme as inhibitors or activators. IC50 value -the concentration of inhibitor which reduces enzyme activity by 50%- was estimated to be 10 mM. Ki constants were also calculated as 6.38 ± 0,70 mM with GSH substrate, and 3.86 ± 0,78 mM with CDNB substrate using the equations of graphs for the inhibitor, and its inhibition type was determined as non-competitive.

Keywords: Drug Resistance, inhibition, glutathione S-transferase

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[1] A. Meister, M.E. Anderson,"Glutathione”, Annu. Rev. Biochem., 1983;52:711-760.
[2] L. Gate, K.D. Tew,"Glutathione S-transferases as emerging targets”, Expert. Opin. Ther. Targets, 2001, 5:477-489.
[3] B. Tozkoparan, S.P. Aytaç,"Kanser kemoterapisinde terapötik hedef olarak glutatyon S-transferazlar”,Hacettepe Üniv. Eczacılık Fak. Dergisi,2007,27:139-164.
[4] K.D. Tew,"Glutathione-associated enzymes in anticancer drug resistance”, Cancer Res., 1994; 54:4313-4320.
[5] J.D. Hayes, D.J. Pulford,"The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance”.Crit. Rev. Biochem. Mol. Biol., 1995, 30:445-600.
[6] R.N. Armstrong,"Structure, catalytic mechanism, and evolution of the glutathione transferases”,Chem. Res. Toxicol.1997, 10:2-18.
[7] T.M. Buetler,D.L. Eaton,"Complementary DNA cloning, messenger RNA expression, and induction of alpha-class glutathione S-transferases in mouse tissues”,Cancer Res., 1992,5:314-318.
[8] U. Tidefelt, A. Elmhorn-Rosenborg, C. Paul, X.Y. Hao, B. Mannervik, L.C. Eriksson,"Expression of glutathione S-transferase-pi as a predictor for treatment results at different stages of acute nonlymphoblastic leukemia”, Cancer Res., 1992, 52:3281-3285.
[9] J.A. Green, L.J. Robertson, A.H. Clark,"Glutathione S-transferase expression in bening and malignant ovarian tumors”, Br. J. Cancer 1993, 68:235-23.
[10] L. Gilbert, L.J. Elwood,M. Merino, S. Masood, R. Barnes, S.M. Steinberg, D.F. Lazarous, L. Pierce, T. d’Angelo, J.A. Moscow,"A pilot study of pi-class glutathione Stransferase expression in breast cancer: correlation with estrogen receptor expression and prognosis in nodenegative breast cancer”, J. Clin. Oncol., 1993,11:49-58.
[11] D.J. Grignon, M. Abdel-Malak, W.C. Mertens, W.A. Sakr, R.R. Shepherd,"Glutathione S-transferase expression in renal cell carcinoma: a new marker of differentiation”, Mod. Pathol., 1994, 7:186-189.
[12] M.L. O’Brien, K.D. Tew,"Glutathione and related enzymes in multidrug resistance”, Eur. J. Cancer, 1996, 6:967-978.
[13] K.D. Tew, S. Dutta, M. Schultz,"Inhibitors of glutathione S-transferases as therapeutic agents”, Adv. Drug Deliv. Rev., 1997, 26:91-104.
[14] D.M. Townsend, K.D. Tew,"The role of glutathione-S-transferase in anti-cancer drug resistance”, Oncogene, 2003, 22:7369-7375.
[15] S. Mahajan, W.M. Atkins,"The chemistry and biology of inhibitors and pro-drugs targeted to glutathione S-transferases”, Cell Mol. Life Sci., 2005, 62:1221-1233.
[16] G. Zhao, X. Wang,"Advance in Antitumor Agents Targeting lutathione- S-Transferase”, Curr. Med. Chem., 2006, 13:1461-1471.
[17] E. Beutler,"Red cell metabolism”, A manual of biochemical methods, Grune and Stratton Inc., 3nd Ed., Orlando, 1984.
[18] P.C. Simons,D.L. Vander Jagt,"Purification of glutathione S-transferases by glutathione-affinity chromatography”,Methods Enzymol., 1981, 77:235-237.
[19] W.H. Habig, W.B.Jakoby,"Assays for differentiation of glutathione Stransferases”. Methods Enzymol., 1981,77:398-405.
[20] M.M.Bradford,"A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding”, Anal. Biochem., 1976,72:248-251.
[21] U.K.Laemmli,"Cleavage of structural proteins during assembly of the head of bacteriophage T4”, Nature, 1970, 227:680-683.
[22] M. Erat, H. Sakiroglu,"The effect of some antineoplastic agents on glutathione S-transferase from human erythrocytes”,J. Enzyme Inhibit. Med. Chem., 2013, 28:711-716.
[23] H. Lineweaver, D.Burk,"The determination of enzyme dissociation constant”, J. American Chem.Soc., 1934,56:658-666.
[24] K.D. Tew,"Glutathione-associated enzymes in anticancer drug resistance”. Cancer Res., 1994, 54:4313-4320.
[25] G. Damia, M. D’Incalci,"Mechanisms of resistance to alkylating agents”,Cytotechnology,1998,27:165-173.
[26] M.L. O’Brien, K.D. Tew,"Glutathione and related enzymes in multidrug resistance”, Eur. J. Cancer, 1996, 32A:967-978.
[27] D.J. Waxman,"Glutathione S-transferases: role in alkylating agent resistance and possible target for modulation chemotherapy”, Cancer Res. 1990, 50:6449-6454.
[28] S. Tsuchida, K. Sato,"Glutathione transferases and cancer”, Crit. Rev. Biochem. Mol. Biol., 1992, 27:337-384.
[29] M. Bredel,"Anticancer drug resistance in primary human brain tumors”, Brain Research Reviews, 2001, 35:161-204.
[30] S. Mahajan, W.M. Atkins,"The chemistry and biology of inhibitors and pro-drugs targeted to glutathione S-transferases”, Cell Mol. Life Sci., 2005, 62:1221-1233.
[31] D. Burg, D.V. Filippov, R. Hermanns, G.A. van der Marel, J.H. van Boomb, G.J. Muldera,"Peptidomimetic glutathione analogues as novel γGT Stable GST inhibitors”. Bioorg. Med. Chem., 2002, 10:195-205.
[32] M.H. Lyttle, M.D. Hocker, H.C. Hui, C.G. Caldwell, D.T. Aaron, A. Engqvist-Goldstein, J.E. Flaggaard, K.E. Bauer,"Isozyme-specific glutathione-S-transferase inhibitors: design and synthesis”, J. Med. Chem., 1994, 37:189-194.
[33] J.J. van Zanden,L. Geraets, H.M. Wortelboer, P.J. van Bladeren,I.M. Rietjens, N.H. Cnubben,"Structural requirements for the flavonoidmediated modulation of glutathione S-transferase P1-1 and GS-X pump activity in MCF7 breast cancer cells”, Biochem. Pharmacol. 2004,67:1607-1617.
[34] Z. Wu, G.S. Minhas, D. Wen, H. Jiang, K. Chen, P. Zimniak, J. Zheng,"Design, synthesis and structure-activity relationships of haloenol lactones: site-directed and isozyme-selective glutathione S-transferase inhibitors”, J. Med. Chem., 2004, 47:3282-3294.
[35] P. Turella, C. Cerella, G. Filomeni, A. Bullo, F. De Maria, L. Ghibelli, M.R. Ciriolo, M. Cianfriglia, M. Mattei, G. Federici, G. Ricci, A.M. Caccuri,"Proapoptotic activity of new glutathione S-transferase inhibitors”, Cancer Res. 2005, 65:3751-3761.
[36] S.A. Mandel, T. Amit, O. Weinreb, L. Reznichenko, M.B.H. Youdim,"Simultaneous Manipulation of Multiple Brain Targets by Green Tea Catechins: A Potential Neuroprotective Strategy for Alzheimer and Parkinson Diseases”, CNS Neuroscience Therapeutics, 2008,14:352-365.
[37] A.G. Dulloo, J. Seydoux, L. Girardier, P. Chantre, J.Vandermander, "Green tea and thermogenesis: interactions between catechinpolyphenols, caffeine and sympathetic activity”,Int. Journal Obesity, 2000,24:252-258.