DOI: https://doi.org/10.61435/jbes.2024.19940

Kinetic features and characterization of liver glutathione transferase in rats exposed to glyphosate

1Department of Biochemistry, College of Biosciences, Federal University of Agriculture, Abeokuta, Nigeria

2Department of Animal Nutrition, College of Animal Science and Livestock Production, Federal University of Agriculture, Abeokuta, Nigeria

3Department of Biochemistry, College of Health Sciences, Osun State University, Osogbo, Nigeria

4 Department of Biochemistry, Faculty of Natural and Applied Sciences, Lead City University, Ibadan, Nigeria

View all affiliations
Received: 22 Jul 2024; Revised: 8 Oct 2024; Accepted: 3 Dec 2024; Available online: 3 Feb 2025; Published: 1 Apr 2025.
Editor(s): H. Hadiyanto
Open Access Copyright (c) 2025 The Author(s). Published by Centre of Biomass and Renewable Energy (CBIORE)
Creative Commons License This work is licensed under a Creative Commons Attribution 4.0 International License.

Citation Format:
Abstract

Glutathione-S-transferases (GSTs) are phase-II metabolizing enzymes which detoxify various compounds through their conjugation reaction. This study characterized and investigated the kinetic properties of liver glutathione-S-transferase in rats exposed to glyphosate. Rats of average weight 200 g were randomly divided into groups (n=4); A, B, C and D. Group A served as control and received distilled water alone while groups B, C and D were exposed to 200, 300 and 400 mg/kg body weight of glyphosate respectively. Three GST model substrates [1, 2-dichloro-4-nitrobenzene (DCNB), paranitrobenzylchloride (pNBCl), and 1-chloro-2, 4-dinitrobenzene (CDNB)] were used to determine the substrate utilization pattern of the GST isozymes in the liver crude homogenate. The liver homogenate was purified by combination of 80 % ammonium sulfate precipitation, gel filtration chromatography on Sephacryl S-200 column and ion exchange chromatography on CM-Sepharose column. Spectrophotometric methods were used to determine the activity of the GSTs and protein concentration. GST isozymes in the liver homogenate were unable to conjugate pNBCl to GSH, DCNB showed slight conjugation while CDNB shows a 2-fold conjugation. The optimum temperature for the induced isozymes of GST A and GST B were 400C and 500C respectively while the optimum pH were 8.0 and 8.5 respectively. In this study, pNBCl proved unsuitable due to its inability to conjugate GSH. Future study may provide additional information about the class of GST isozyme induced.

Note: This article has supplementary file(s).

Fulltext View|Download |  common.other
Untitled
Subject
Type Other
  Download (B)    Indexing metadata
Keywords: Glutathione transferase (GST); Glyphosate; Metal ions; Thiol reagents

Article Metrics:

Article Info
Section: Research Articles
Language : EN
Recent articles
Physico-Chemical Properties and Bacteriological Analysis of Mangrove Water from Mangrove Forest of Gbaramatu Kingdom Delta, Nigeria Treatment of Cheese Whey and Bioelectricity Generation in MFCs as Substitute Source of Energy in Wastewater Kinetic features and characterization of liver glutathione transferase in rats exposed to glyphosate More recent articles
  1. Adewale, I. O., and Afolayan, A. (2005). Organ distribution and kinetics of Glutathione transferase from African river prawn, Macrobrachium vollenhovenii (Herklots). Aquatic Toxicology, 71(2), 193-202. https://doi.org/10.1016/j.aquatox.2004.11.005
  2. Adewale, I. O., and Afolayan, A. (2006). Studies on glutathione transferase from grasshopper (Zonocerus variegatus). Pesticide Biochemistry and Physiology, 85(1), 52-59. https://doi.org/10.1016/j.pestbp.2005.10.004
  3. Agunbiade, J. O., Adewale, I. O., and Afolayan, A. (2014). Kinetic analysis of glutathione transferase from rats exposed to sub-lethal and lethal concentrations of lead acetate. Ife Journal of Science, 16(3):417-423
  4. Ahmed, S., Sohail, A., Khatoon, S., Khan, S., and Saifullah, M. K. (2017). Partial purification and characterization of glutathione S-transferase from the somatic tissue of Gastrothylax crumenifer (Trematoda: Digenea). Veterinary World, 10(12), 1493-1500. https://doi.org/10.14202/vetworld.2017.1493-1500
  5. Akkemik, E., Taser, P., Bayindir, A., Budak, H., and Ciftci, M. (2012). Purification and characterization of glutathione S-transferase from Turkey liver and inhibition effects of some metal ions on enzyme activity. Environmental Toxicology and Pharmacology, 34(3), 888-894. https://doi.org/10.1016/j.etap.2012.08.010
  6. Aksoy, M., Ozaslan, M. S., and Kufrevioglu, O. I. (2016). Purification of glutathione S-transferase from Van Lake fish (Chalcalburnus tarichii Pallas) muscle and investigation of some metal ions effect on enzyme activity. Journal of Enzyme Inhibition and Medicinal Chemistry, 31(4), 546–550. https://doi.org/10.3109/14756366.2015.1046063
  7. Anushia, C., Kumar, P. S., and Karthikeyan, P. (2012). Heavy metal induced enzyme response in Tilapia mossambicus. International Journal of Pharmaceutical Research and Bio-Science, 1:371–385
  8. Basantani, M., and Srivastava, A. (2007). Plant glutathione transferases — a decade falls short. Canadian Journal of Botany, 85(5), 443-456. https://doi.org/10.1139/b07-033
  9. Basha, P., and Rani, A. (2003). Cadmium-induced antioxidant defense mechanism in freshwater teleost Oreochromis mossambicus (Tilapia). Ecotoxicology and Environmental Safety, 56(2), 218-221. https://doi.org/10.1016/s0147-6513(03)00028-9
  10. Biswas, S., Chida, A. S., and Rahman, I. (2006). Redox modifications of protein–thiols: Emerging roles in cell signaling. Biochemical Pharmacology, 71(5), 551–564. https://doi.org/10.1016/j.bcp.2005.10.044
  11. Bradford, M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72(1-2), 248-254. https://doi.org/10.1006/abio.1976.9999
  12. Cerdeira, A. L., and Duke, S. O. (2006). The current status and environmental impacts of glyphosate‐resistant crops. Journal of Environmental Quality, 35(5), 1633-1658. https://doi.org/10.2134/jeq2005.0378
  13. Comakli, V., Ciftci, M., and Kufrevioglu, O. I. (2012). Effects of some metal ions on rainbow trout erythrocytes glutathione S-transferase enzyme: Anin vitrostudy. Journal of Enzyme Inhibition and Medicinal Chemistry, 28(6), 1261-1266. https://doi.org/10.3109/14756366.2012.729829
  14. Çomaklı, V., Kuzu, M., and Demirdağ, R. (2015). Characterization and purification of Glutathione S‐transferase from the liver and Gill tissues of Ağrı Balık lake trout Salmo trutta labrax and the effects of heavy metal ions on its activity. Journal of Aquatic Animal Health, 27(3), 145-151. https://doi.org/10.1080/08997659.2015.1032441
  15. Daggett, D. A., Oberley, T. D., Nelson, S. A., Wright, L. S., Kornguth, S. E., and Siegel, F. L. (1998). Effects of lead on rat kidney and liver: GST expression and oxidative stress. Toxicology, 128(3), 191-206. https://doi.org/10.1016/s0300-483x(98)00080-8
  16. Demirdag, R., Yerlikaya, E., Kufrevioglu, O. I., and Gundogdu, C. (2012). Purification of glutathiones-transferase isoenzymes from tumour and nontumour human stomach and inhibitory effects of some heavy metals on enzymes activities. Journal of Enzyme Inhibition and Medicinal Chemistry, 28(5), 911-915. https://doi.org/10.3109/14756366.2012.694878
  17. Dixon, D. P., and Edwards, R. (2009). Selective binding of Glutathione conjugates of fatty acid derivatives by plant Glutathione transferases. Journal of Biological Chemistry, 284(32), 21249-21256. https://doi.org/10.1074/jbc.m109.020107
  18. Dobritzsch, D., Grancharov, K., Hermsen, C., Krauss, G., and Schaumlöffel, D. (2020). Inhibitory effect of metals on animal and plant glutathione transferases. Journal of Trace Elements in Medicine and Biology, 57, 48-56. https://doi.org/10.1016/j.jtemb.2019.09.007
  19. Duke, S. O., Baerson, S. R., and Rimando, A. M. (2003). Glyphosate. Encyclopedia of Agrochemicals. https://doi.org/10.1002/047126363x.agr119
  20. Duke, S. O., and Powles, S. B. (2008). Glyphosate: A once‐in‐a‐century herbicide. Pest Management Science, 64(4), 319-325. https://doi.org/10.1002/ps.1518
  21. Fjellstedt, T. A., Allen, R. H., Duncan, B. K., and Jakoby, W. B. (1973). Enzymatic conjugation of epoxides with Glutathione. Journal of Biological Chemistry, 248(10), 3702-3707. https://doi.org/10.1016/s0021-9258(19)43983-5
  22. Gadagbui, B. K., and James, M. O. (2000). Activities of affinity-isolated glutathione S-transferase (GST) from channel catfish whole intestine. Aquatic Toxicology, 49(1-2), 27-37. https://doi.org/10.1016/s0166-445x(99)00073-9
  23. Garcia-Viloca, M., Gao, J., Karplus, M., and Truhlar, D. G. (2004). How enzymes work: Analysis by modern rate theory and computer simulations. Science, 303(5655), 186-195. https://doi.org/10.1126/science.1088172
  24. Glisic, B., Mihaljevic, I., Popovic, M., Zaja, R., Loncar, J., Fent, K., Kovacevic, R., and Smital, T. (2015). Characterization of glutathione-S-transferases in zebrafish (Danio rerio). Aquatic Toxicology, 158, 50-62. https://doi.org/10.1016/j.aquatox.2014.10.013
  25. Gronwald, J. W., and Plaisance, K. L. (1998). Isolation and characterization of glutathiones-transferase Isozymes from Sorghum1. Plant Physiology, 117(3), 877-892. https://doi.org/10.1104/pp.117.3.877
  26. Guddewar, M. B., and Dauterman, W. C. (1979). Purification and properties of a glutathione-S-transferase from corn which conjugates S-triazine herbicides. Phytochemistry, 18(5), 735-740. https://doi.org/10.1016/0031-9422(79)80005-9
  27. Habig, W. H., Pabst, M. J., and Jakoby, W. B. (1974). Glutathione S-transferases. Journal of Biological Chemistry, 249(22), 7130-7139. https://doi.org/10.1016/s0021-9258(19)42083-8
  28. Hayes, J. D., and Pulford, D. J. (1995). The glutathione S-transferase supergene family: Regulation of GST and the contribution of the lsoenzymes to cancer Chemoprotection and drug resistance part II. Critical Reviews in Biochemistry and Molecular Biology, 30(6), 521-600. https://doi.org/10.3109/10409239509083492
  29. Hayes, J. D., Flanagan, J. U., and Jowsey, I. R. (2005). Glutathione transferases. Annual Review of Pharmacology and Toxicology, 45(1), 51-88. https://doi.org/10.1146/annurev.pharmtox.45.120403.095857
  30. Huang, S., and Han, Z. (2007). Mechanisms for multiple resistances in field populations of common cutworm, Spodoptera litura (Fabricius) in China. Pesticide Biochemistry and Physiology, 87(1), 14-22. https://doi.org/10.1016/j.pestbp.2006.05.002
  31. Ketterer, B. (1971). Amino-azo-dye-binding protein in the soluble cytoplasm of the rat liver. Biochemical Journal, 122, 53–54. https://doi.org/10.1042/bj1220053pb
  32. Koshland, D. E. (1958). Application of a theory of enzyme specificity to protein synthesis. Proceedings of the National Academy of Sciences, 44(2), 98–104
  33. Ladner, J. E., Parsons, J. F., Rife, C. L., Gilliland, G. L., and Armstrong, R. N. (2003). Parallel evolutionary pathways for Glutathione Transferases: structure and mechanism of the mitochondrial class kappa enzyme rgstk1-1. Biochemistry, 43(2), 352-361. https://doi.org/10.1021/bi035832z
  34. Louhimies, S. (2002). Directive 86/609/EEC on the protection of animals used for experimental and other scientific purposes. ATLA 30:217–219
  35. McLellan, L. I., and Wolf, C. (1999). Glutathione and glutathione-dependent enzymes in cancer drug resistance. Drug Resistance Updates, 2(3), 153-164. https://doi.org/10.1054/drup.1999.0083
  36. Meyer, D. J., Coles, B., Pemble, S. E., Gilmore, K. S., Fraser, G. M., and Ketterer, B. (1991). Theta, a new class of glutathione transferases purified from rat and man. Biochemical Journal, 274(2), 409-414. https://doi.org/10.1042/bj2740409
  37. Morgenstern, R. (2024). Kinetic behavior of glutathione transferases: Understanding cellular protection from reactive intermediates. Biomolecules, 14(6), 641. https://doi.org/10.3390/biom14060641
  38. Nagy, P. (2013). Kinetics and mechanisms of thiol–disulfide exchange covering direct substitution and thiol oxidation-mediated pathways. Antioxidants & Redox Signaling, 18(13), 1623-1641
  39. Ojopagogo, Y. A., Adewale, I. O., Adeyemi, J. A., and Afolayan, A. (2015). Some novel features of glutathione transferase from juvenile catfish (Clarias gariepinus) exposed to lindane-contaminated water. Perspectives in Science, 4, 62-65. https://doi.org/10.1016/j.pisc.2014.12.010
  40. Özaslan, M. S., Demir, Y., Aksoy, M., Küfrevioğlu, Ö. I., and Beydemir, Ş. (2018). Inhibition effects of pesticides on glutathione‐s‐transferase enzyme activity of van lake fish liver. Journal of Biochemical and Molecular Toxicology, 32(9). https://doi.org/10.1002/jbt.22196
  41. Pandey, T., Singh, S. K., Chhetri, G., Tripathi, T., and Singh, A. K. (2015). Characterization of a highly pH stable CHI-class Glutathione S-transferase from Synechocystis PCC 6803. PLOS ONE, 10(5), e0126811. https://doi.org/10.1371/journal.pone.0126811
  42. Pauling, L. (1946). Molecular architecture and biological reactions. Chemical & Engineering News Archive, 24(10), 1375-1377. https://doi.org/10.1021/cen-v024n010.p1375
  43. Ranson, H., Rossiter, L., Ortelli, F., Jensen, B., Wand, X., Roth, C. W., Collins, F. H., and Hemingway, J. (2001). Identification of a novel class of insect glutathione S-transferases involved in resistance to DDT in the malaria vector anopheles gambiae. Biochemical Journal, 359(2), 295. https://doi.org/10.1042/0264-6021:3590295
  44. Ricci, G., Caccuri, A.M., Lo Bello, M., Parker, M.W., Nuccetelli, M., Turella, P., Stella, L., Di Iorio, E.E., Federici, G. (2003). Glutathione transferase P1-1: Self-preservation of an anti-cancer enzyme. Biochemical Journal, 376, 71–76. https://doi.org/10.1042/bj20030860
  45. Rubino, F. (2015). Toxicity of glutathione-binding metals: A review of targets and mechanisms. Toxics, 3(1), 20-62. https://doi.org/10.3390/toxics3010020
  46. Sau, A., Pellizzari Tregno, F., Valentino, F., Federici, G., and Caccuri, A. M. (2010). Glutathione transferases and development of new principles to overcome drug resistance. Archives of Biochemistry and Biophysics, 500(2), 116-122. https://doi.org/10.1016/j.abb.2010.05.012
  47. Sheehan, D., Meade, G., Foley, V. M., and Dowd, C. A. (2001). Structure, function and evolution of glutathione transferases: Implications for classification of non-mammalian members of an ancient enzyme superfamily. Biochemical Journal, 360(1), 1. https://doi.org/10.1042/0264-6021:3600001
  48. Shimoji, M., Aniya, Y., and Morgenstern, R. (2007). Activation of microsomal glutathione transferase 1 in toxicology. In Y. C. Awasthi (Ed.), Toxicology of glutathione transferases (pp. 293–319). Boca Raton, FL: CRC Press/Taylor & Francis Group
  49. Singhal, S. S., Singh, S. P., Singhal, P., Horne, D., Singhal, J., and Awasthi, S. (2015). Antioxidant role of glutathione S-transferases: 4-Hydroxynonenal, a key molecule in stress-mediated signaling. Toxicology and Applied Pharmacology, 289(3), 361-370. https://doi.org/10.1016/j.taap.2015.10.006
  50. Spearman, M. E., Prough, R. A., Estabrook, R. W., Falck, J., Manna, S., Leibman, K. C., Murphy, R. C., and Capdevila, J. (1985). Novel glutathione conjugates formed from epoxyeicosatrienoic acids (EETs). Archives of Biochemistry and Biophysics, 242(1), 225-230. https://doi.org/10.1016/0003-9861(85)90496-5
  51. Thomson, R. E., Bigley, A. L., Foster, J. R., Jowsey, I. R., Elcombe, C. R., Orton, T. C., and Hayes, J. D. (2004). Tissue-specific expression and subcellular distribution of murine Glutathione S-transferase class kappa. Journal of Histochemistry & Cytochemistry, 52(5), 653-662. https://doi.org/10.1177/002215540405200509
  52. Turkan, F., Sakiroglu, H., and Balci, N. (2014). Purification and characterization of the glutathione-S-transferase enzyme from the fruit fruit (Laurocerasus officinalis Roem.) and inhibition effects of enzyme activity. Mus Alparslan University Journal of Science 2(2):280-288. https://dergipark.org.tr/en/pub/msufbd/issue/19659/370826
  53. Turkanoglu, A. (2007). Human serum arylesterase and glutathione s-transferase activities in patients with ischemic stroke compared to healthy controls. Master of Science Dissertation, Middle East Technical University. https://open.metu.edu.tr/handle/11511/17408
  54. Vašková, J., Kočan, L., Vaško, L., & Perjési, P. (2023). Glutathione-related enzymes and proteins: A review. Molecules, 28(3), 1447. https://doi.org/10.3390/molecules28031447
  55. Wan, H., Zhan, S., Xia, X., Xu, P., You, H., Jin, B. R., and Li, J. (2016). Identification and functional characterization of an epsilon glutathione S-transferase from the beet armyworm (Spodoptera exigua). Pesticide Biochemistry and Physiology, 132, 81-88. https://doi.org/10.1016/j.pestbp.2015.09.009

Last update:

No citation recorded.

Last update:

No citation recorded.

echo '
https://journals.aesop-planning.eu/ https://jurnal.pgsd.unipol.ac.id/ https://superbilisim.com.tr/ https://journal.pubalaic.org/
';