Advances in Biochemistry
Volume 2, Issue 5, October 2014, Pages: 71-75
Received: Sep. 7, 2014;
Accepted: Sep. 19, 2014;
Published: Oct. 30, 2014
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Ghada Al-Amro, Dept. of Biochemistry, King Saud University, Riyadh 11495, Saudi Arabia
Mohammad Ali Qorban, Dept. of Biochemistry, King Saud University, Riyadh 11495, Saudi Arabia
Samina Hyder Haq, Dept. of Biochemistry, King Saud University, Riyadh 11495, Saudi Arabia
Glutathione S-transferases (GST) are a group of multifunctional ubiquitous enzymes widely present in animals and plants, which catalysis the conjugation of glutathione to different exogenous and endogenous electrophilic compounds. This study was carried out to characterize the purified GST enzyme from camel liver tissues and to investigate the in-vitro inhibitory effect of the flavonoid quercetin by measuring S-2,4-dinitrophenyl glutathione (DNP-GSH) formation from 1-chloro-2,4-dinitrobenzene (CDNB) and reduced glutathione(GSH) as substrates. The Km values for reduced GSH and CDNB were found to be 0.08438 and 0.6827 mM while Vmax values were 6.935 and 15.599 mM/min respectively. The IC50 was determined to be 1.8 mM. The inhibition constant (Ki) was estimated to be 1.91 mM at 0.5 mM and 1.76 mM at 2 mM. The mean inhibition constant (Ki) was estimated to be 1.835±0.075mM which revealed an uncompetitive profile and indicated quercetin as a weak inhibitor with the varied concentration of CDNB and fixed concentration of reduced GSH as a substrate.
Mohammad Ali Qorban,
Samina Hyder Haq,
In-Vitro Inhibition of Camel Hepatic Glutathione Transferase by Quercetin, Advances in Biochemistry.
Vol. 2, No. 5,
2014, pp. 71-75.
Sheehan D, Meade G, Foley VM, Dowd CA. Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochem J. 2001;360(Pt 1):1-16.
Liu D, Liu Y, Rao J, Wang G, Li H, Ge F, et al. [Overexpression of the glutathione S-transferase gene from Pyrus pyrifolia fruit improves tolerance to abiotic stress in transgenic tobacco plants]. Mol Biol (Mosk). 2013;47(4):591-601.
Park HJ LK, Cho SH, Kong KH. Functional studies of cysteine residues in human glutathion-S-transferase P1-1 by site-directed mutagenesis. Bull Korean Chem. 2001;22:77-82.
Morgenstern R, DePierre JW, Jornvall H. Microsomal glutathione transferase. Primary structure. J Biol Chem. 1985;260(26):13976-83.
Armstrong RN. Glutathione S-transferases: reaction mechanism, structure, and function. Chem Res Toxicol. 1991;4(2):131-40.
Josephy PD. Genetic variations in human glutathione transferase enzymes: significance for pharmacology and toxicology. Hum Genomics Proteomics. 2010;2010:876940.
Hayes JD, Pulford DJ. 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(6):445-600.
Adler V, Yin Z, Fuchs SY, Benezra M, Rosario L, Tew KD, et al. Regulation of JNK signaling by GSTp. Embo j. 1999;18(5):1321-34.
Cho SG, Lee YH, Park HS, Ryoo K, Kang KW, Park J, et al. Glutathione S-transferase mu modulates the stress-activated signals by suppressing apoptosis signal-regulating kinase 1. J Biol Chem. 2001;276(16):12749-55.
Raza H, John A, Lakhani MS, Ahmed I, Montague W. Multiplicity and tissue specific expression of camel cytochrome P450(s). Comp Biochem Physiol C Pharmacol Toxicol Endocrinol. 1998;121(1-3):205-11.
Moskaug JO, Carlsen H, Myhrstad M, Blomhoff R. Molecular imaging of the biological effects of quercetin and quercetin-rich foods. Mech Ageing Dev. 2004;125(4):315-24.
Hertog MG, Kromhout D, Aravanis C, Blackburn H, Buzina R, Fidanza F, et al. Flavonoid intake and long-term risk of coronary heart disease and cancer in the seven countries study. Arch Intern Med. 1995;155(4):381-6.
Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974;249(22):7130-9.
Hunaiti AA, Abukhalaf IK. A rapid purification procedure for camel kidney glutathione S-transferase. Prep Biochem. 1987;17(3):239-59.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-75.
van Zanden JJ, Ben Hamman O, van Iersel ML, Boeren S, Cnubben NH, Lo Bello M, et al. Inhibition of human glutathione S-transferase P1-1 by the flavonoid quercetin. Chem Biol Interact. 2003;145(2):139-48.
Merlos M, Sanchez RM, Camarasa J, Adzet T. Flavonoids as inhibitors of rat liver cytosolic glutathione S-transferase. Experientia. 1991;47(6):616-9.
Sahu SC, Gray GC. Pro-oxidant activity of flavonoids: effects on glutathione and glutathione S-transferase in isolated rat liver nuclei. Cancer Lett. 1996;104(2):193-6.
Wiegand H, Boesch-Saadatmandi C, Regos I, Treutter D, Wolffram S, Rimbach G. Effects of quercetin and catechin on hepatic glutathione-S transferase (GST), NAD(P)H quinone oxidoreductase 1 (NQO1), and antioxidant enzyme activity levels in rats. Nutr Cancer. 2009;61(5):717-22.
Bousova I, Skalova L. Inhibition and induction of glutathione S-transferases by flavonoids: possible pharmacological and toxicological consequences. Drug Metab Rev. 2012;44(4):267-86.