Study of Thioredoxin Isolated from Human Blood Serum

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Luay A. Al-Helaly
Amera A. Hamdon

Abstract

The research included the separation of thioredoxin (Trx) from human blood serum using different biochemical techniques. One proteinous peak had been isolated by gel filtration using sephadex )G-50) that produced by ammonium sulphate precipitation(65%) after dialysis.  It was noted that there are two peaks of Trx when using ion exchange-type DEAE-cellulose for the product of gel filtration column.


The approximately molecular weight of the isolated Trx using gel filtration chromatography (G-50) and electrophoresis technique type SDS-PAGE were (12305 + 190) Dalton and 12461 Dalton respectively .


The results showed that inhibited and activated effects of drugs on Trx working. The results showed that inhibited to 44.59% for paracetamol, dexathamazone, diphenhydramine-HCl, theophlline anhydrous,  pseudoephedrine, ceramide and caffeine anhydrous, but metronidazole and chlorpheniramine malate have activated effect to 110.8% and 21.62 respectively. 

Article Details

How to Cite
Luay A. Al-Helaly, & Amera A. Hamdon. (2023). Study of Thioredoxin Isolated from Human Blood Serum. Tikrit Journal of Pure Science, 20(3), 79–88. https://doi.org/10.25130/tjps.v20i3.1190
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References

1-Williams, JS.C.H. (1995). "Chemistry and Biochemistry of Flavonezymes" (Muller, F., ed). CRC Press. Baco Raton. Pp.121-211.

2-Hashemy,S.I.(2011). The human thioredoxin system: modifications and clinical applications. Iranian Journal of Basic Medical Sciences. 14(3),191-204

3- Hashemy, S.; Ungerstedt, J.; Zahedi Avval, F.; Holmgren, A. (2006). Motexafin gadolinium, a tumor-selective drug targeting thioredoxin reductase and ribonucleotide reductase. J. Biol. Chem. 281 (16): 10691–7.

4- Mustacich, D.; Powis, G.(2000). Thioredoxin reductase. Biochem. 267, 6102.

5-Arner, E.S.; Holmgren, A. (2006). The thioredoxin system in cancer. Semin Caner Biol. 16(6), 420-6.

6- Watson, W. H., Yang, X., Choi, Y.E., Jones, D.P., and Kehrer, J. P.(2004). Thioredoxin and its role in toxicology. Toxicol Sci.78,3-14.

7- Holmgren ,A.(1979). Thioredoxin catalyzes the reduction of insulin disulfides by dithiothreitol and dihydrolipoamide. J. Biol. Chem. 254(19), 9626-9632.

8- Schacterle, G. R.; Pollack, R. L. (1973). A simplified method for the quantitative assay of small amount of protein in biological material. Anal. Biochem. 51, 654-655.

9- Maggioli, G.;Piacenza,L.; Carambula, B.; Carmona, C. (2004). Purification characterization and immunolo calization of a thioredoxin reductase from adult Fasciola Hepatica. J .Paraitol.,90(2), 205-211.

10- Plummer, T.D. (1978). An introduction to practical biochemistry. 2nd. Ed., McGraw-Hill Book Comp., U.K.

11-Holmgren, A.(1977). Bovine thioredoxin system. Purification of thioredoxin reductase from calf liver and thymus and studies of its function in disulfide reduction. J. Biol. Chem., 252, 4600-4606.

12- Robyt, F.J.; White, J.B. (2001). "Biochemical Techniques, Theory and Practice", books/cole publishing company, Monterey, California.

13- Burtis, C. A.; Ashwood, E. R.; Bruns, D.E.(2012). "Tietz Textbook of Clinical Chemistry and Molecular Diagnostics". By Saunders, an imprint of Elsevier Inc. USA. pp.356, 368.

14- Murray, R. K.; Bender, D. A.; Botham, K. M.; Kennelly, P. J.; Rodwell, V. W. (2009)." Harper's Illustrated Biochemistry". 28 ed. The McGraw-Hill Companies, Chapter 8.

15- Gromer, S.; Arscott, L.D.; Williams, C.H.J.; Schirmer, R.H. (1998). Human placenta thioredoxin Reductase. Isolation of the selenoenzymes, steady state kinetics and inhibition by therapeutic gold compounds. J. Biol. Chem., 273, 20096-20101.

16- Rackham, O.; Sheanwood, A. M.; Thyer,R.;Mc Namara, E.;Davies,SM.; Callus,BA.; Miranda-Vizuete,A. (2011). Substrate and inhibitor specificities differ between human cytosolic and mitochondrial reductase : Implication for development of specific inhibitors. Radic .Biol. Med. 50(6), 689-99.

17- Wang, Q., Hou, Y., Qu, J., Hong, Y., Lin, Y., Han, X.(2013) Molecular cloning, expression, purification and characterization of thioredoxin from Antarctic sea-ice bacteria Pseudoalteromonas sp. AN178. Mol. Biol. Rep. 40(12):6587-91.

18- Harris C., Hansen, J.M.(2012). Oxidative stress, thiols, and redox profiles. Methods Mol Biol. 889:325-46.

19- Harvey, R. A.; Ferrier, D. R. (2014). " Lippincott’s Illustrated Reviews: Biochemistry" 5th ed. Lippincott Williams and Wilkins, a Wolters Kluwer business, China. P.20,

20- Park, C.I., Jung, J.H., Shim, W.J., Kim, J.W., Kim, E.G., Jeong, J.M., Kim, D.H. (2012). Molecular characterization, expression, and functional analysis of two thioredoxins in the black rockfish (Sebastes schlegelii). Fish Shellfish Immunol. 32(5):808-15.

21- Horecká, T., Perecko, D, Kutejová, E, Muchová, K, Kollárová, M. (1996). Purification and partial characterization of two thioredoxins from Streptomyces aureofaciens. Biochem Mol. Biol. Int. 40(3):497-505.

22- Arsova, B., Hoja, U., Wimmelbacher, M., Greiner, E., Ustun, S., Melzer, M., Petersen, K., Lein, W. and Bornke, F. (2010) Plastidial thioredoxin z interacts with two fructokinaselike proteins in a thioldependent manner: evidence for an essential role in chloroplast development in Arabidopsis and Nicotiana benthamiana, Plant Cell, www.plantcell.org/cgi/doi/10.1105/tpc.110.2 20512.

23- Hall, M., Cabana, A.M., Akerlund, H., Florencio, F.J., Schröder, W.P., Lindahl, M. and Kieselbach, T. (2010) Thioredoxin 33 targets of the plant chloroplast lumen and their implications for plastid function. Proteomics, 10: 987–1001.

24- Serrato, A.J. and Cejudo, F.J. (2003) Type-h thioredoxins accumulate in the nucleus of developing wheat seed tissues suffering oxidative stress. Planta, 217: 392–399.

25- Ben Bacha, A.G., Mejdoub, H.(2012). Purification and biochemical characterization of an organic-solvent-tolerant thioredoxin from dromedary pancreas. Protein J. 31(1):1-7.

26- Windle ,H. J., A´ ine F., De´irdre, N. E., Kelleher, D.(2000). The thioredoxin system of Helicobacter pylori . J. Biol. Chem. 275(7):5081–5089.

27- Dai, Ch., Wang, M.(2011). Isolation and characterization of thioredoxin and NADPH-dependent thioredoxin reductase from tomato (Solanum lycopersicum). BMB reports 44(10): 692-697.

28- Horak, F.; Zieglmayer, P.; Zieglmayer, R.; Lemell, P.; Yao, R.; Staudinger, H.; Danzig, M. (2009). A placebo-controlled study of the nasal decongestant effect of phenylephrine and pseudoephedrine in the Vienna Challenge Chamber". Annals of Allergy, Asthma and Immunology 102 (2), 116.

29-Nehlig, A.; Daval, J. L.; Debry, G. (1992). Caffeine and the central nervous system: mechanisms of action, biochemical, metabolic and psychostimulant effects. Brain Research Reviews. 17 (2), 139 –170.

30- Barnes, P.J.; Pauwels, R.A. (1994) . Theophylline in the management of a sthma: time for reappraisal. Eur. Respir J. 7,579-591.

31- Sagara, I.M.; Ozak, T.; Morioka, E.; Wada, K.; Hondia, K.; Hori, A.; Matsuya, Y. (2012). Induced of prolonged continous slow wave sleep by blocking cerebral H1-histamine receptors in rats. Br. J. Pharmacol. 165(1), 167-182.

32- You, N.; Tao, K.; Zhao, W.; Bao, P.; Li, R.; You, H.; Zhang, M. (2010). Diphenhydramine modulates cytokins and induced apoptosis in experimental acute pancreatitis. Methods Find Exp. Clin. Pharmacol. 32(5), 3011-8.

33- Aloorkar, N.H.; Bhatia, M.S. (2011). Rapidly disintegrating tabletes of metoclopramide-HCl using novel chemically modified cellulose. Ars. Pharm. 52(3), 26-34.

34- Lustar, I.; Friedland, I. R.; Jafri, H.S.; Wubbel, L.; Ahmad, A.; Trujillo, M.; McCoig, C.C., and McCraken, G.H.(2003). Factors influencing the anti-inflammatory effects of Dexamethasone therapy in experimental pneumococcal meningitis. J. Antimicrob. Chemotherapy., 52, 651-655.

35- Leitsch, D. , Kolarich, D., Duchene, M. (2010). The Flavin inhibitor diphenyleneiodonium renders Trichomonas Vaginalis resistant to metronidazole, inhibits thioredoxin reductase and Flavin reductase, and shuts off hydrogenosomal enzymatic pathways . Mol. Biochem. Parasitology. 171,1:17-2

36- Hua jan, Y., Heck, D. E., Dragomir, A. , Gardner , C. R., Laskin , Debra L. (2014). Acetaminophen reactive intermediates target hepatic thioredoxin reductase .Chem. Res. Toxicol. 4;27(5):882-94.

37- Sreekumar, P. G., Ding, Y., Hinton, D R.(2009). Regulation of thioredoxin by ceramide in retinal pigment epithelial cells. Exp. Eye Res. 88(3)410-417.

38- Asami, K., Inagaki, A., Imura, T., Sekiguchi S., Fujimori K.(2013). Thioredoxin-1 Attenuates early graft loss after Intraportal Islet transplantation in Mice. PLoS ONE 8(8): e70259.

39- Hoshino, T., Nakamura H., Okamoto, M., Kato, S., Araya, S. (2003) Redoxactive protein thioredoxin prevents proinflammatory cytokine- or bleomycin induced lung injury. Am. J. Respir. Crit. Care Med. 168: 1075–1083.

40- Ueda, S., Nakamura, T., Yamada, A., Teratani, A., Matsui, N.(2006). Recombinant human thioredoxin suppresses lipopolysaccharide-induced bronchoalveolar neutrophil infiltration in rat. Life Sci. 79: 1170–1177