Inhibition of Partially Purified Xanthine Oxidase Activity from renal failure patients by Theophylline and Metronidazole
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Abstract
Xanthine oxidase (XO) was purified from sera of patients with renal failure by using ammonium sulfate and dialysis with a specific activities 2.1×10-3 and 5.6×10-3 unit/mg protein respectively. By using anion exchange chromatography techniques, One protein peak of XO activity was obtained with specific activity 78×10-3unit/mg protein and with purification fold of 134.02 compared to crude enzyme. One isoenzyme was obtained having a molecular weight of 255344 Dalton approximately.
Inhibition of purified XO was studied by deferent concentrations of Theophylline and Metronidazole drugs. The maximal inhibitory effect was exhibited with 70 and 80µM Theophylline and 16 and 18 mM metronidazole.
The inhibition type of XO by Theophylline was competitive. Vmax value was 0.0416 unit/ml, and Km value without inhibitor was 0.833 mM, however values were 2 and 2.85 mM, and inhibition constant Ki values were 29.57 and 23.67 mM respectively with 70 and 80µM of Theophylline.
The inhibition type of XO by Metronidazole was non competitive. Km value 0.833 mM and Vmax value without inhibitor was 0.0416 unit/ml. However, Vmax values were 0.027 and 0.023 unit/ml and inhibition constant Ki values were 17 and 19mM by using 16 and 18 mM of Metronidazole respectively.
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References
1. Soni J.P., Parmar D.R. and Sen D.J., Internationale
Pharmaceutica Sciencia, 1(1) : 107-115 (2011).
2. Broges F., Fernandes E. and Roleiva F., Current
Med. Chem., 9(2): 195-217(2002).
3. Linder N., Marthein E., Landen M., Lauhimo J.,
Nordling S. and Haglund C., Eur. J. Cancer,
45(4):648-55 (2009).
4. Jennifer G.D., Rjashree R., Linda B.S. and Anne
M.M., Am. J. Physiol. Heart. Cire. Physiol., 289:
H1518-H1518 (2005).
5. Heunks L.M., Vina J., van Herwaarder C.L.,
Folgering H.T., Gimeno A. and Dekhuijzen P.N., Am
. J . physiol ., 277: 1697-1704 (1999). .
6. Ichinose M., Sugiura H., Yamagata S., Koarai A.,
Tomaki M., Ogawa H., Komaki K., Barnes P.J.,
Shirato K. and Hattori T., Eur. Respir. J., 22: 457-
461 (2003).
7. Pacher P., Nerorozhkin A. and Szabo C.,
Pharmacia. Rev., 58: 87-114 (2006). (IVSL).
8. Chung H.Y., Baek B.S., Song S.H., Kim M.S.,
Huh J.I., Shim K.H., Kim K.W. and Lee K.H.,
20(3): 127-140 (1997). (IVSL)
9. Desco M.C., Asensi M., Marquez R., Martanez-
Valls J., Vento M., Pallard F.V., Sastre J. and Viaa J.,
Diab., 51: 1118-1124 (2002).
10. Al-Leheebi N.I., AL-Abbasy O.Y. and ALSaadon
M.B., Raf.J.Sci.,19(2):48-55(2008).
11. Engberding N., Spiekermann S., Schaefer A.,
Heineke A., Wiencke A., Muller M., Fuchs M.,
Hilfiker-Kleiner D., Hornig B., Drexeler H. and
Landmesser U., Circ. Res., 110: 2175-2179 (2004)..
12. Stull L.B., Leppo M.K., Szweda L., Gao W.D.
and Marban E., Circ. Res., 95: 1005-1011 (2004)..
13. Zhang D.J., Eiswik R.R., Miller G. and Bialy J.L.,
Clin. Chem., 44:1325-1333 (1998).
14. Wootton I.D.P., “Microanalysis in medical
biochemistry”. 5th ed., Churchill Livingstone,
Edinburgh, pp.156-159 (1974)..
15. Ackermann E. and Brill A.S. “Xanthine oxidase
activity, in method of enzymatic analysis”. 2nd .,
Bergameyer , H.U., Academic Press Inc., USA. p521,
(1974).
16. Laemmli UK., Nature, 227:680–685 (1970).
17. Befani O., Grippa E., Saso L., Turini P. and
Mondovi B., Inflamm. Res., 50: 136-137 (2001).
18. Lelas F.B., Raf .J. Sci., 23(4): 70-82 (2012).
19. Siddiqi NJ., Saxena JK., Tripathi LM., Dutta GP.
and Pandey VC., Journal of Parasitic Diseases,
20(2): 155-158 (1996).
20. Maia L. and Mira L., Arch. Biochem. Biophys.,
400 (1):48-53 (2002).
21. Hille, R., Arch. Biochem.Biophys., 443 (1):107-
16 (2005).
22. Moriwaki Y., Yamamoto T., Suda M., Nasako Y.,
Takahashi S., Agbedana O.E., Hada T. and
Higashino K., Biochimica. Biophysica. acta,
1164: 327-330 (1993).
23. Carpani G., Racchi M., Ghezzi P., Terao M. and
Garattini E., Archives of Biochemistry and
Biophysics, 279(2): 237–241 (1990).
24. Xin Y., Yang H., Xia X., Zhang L., Zhang Y.,
Cheng C. and Wang W., Process Biochemistry, 47:
1539-1544 (2012). (IVSL).
25. Elion G.B., Callahan S., Nathan H., Bieber S.,
Raundles W. and Hitching G.H., Biochem.
Pharmacol., 12:85-93 (1963).
26. Hille R. and Stewart RC., J. Biol. Chem.,
259(3):1570–1576 (1984).
27. Jandal J.M., "Fondamental of Biochemistry".
Tikrit University. 1sted ., p276 (2006)..
28. Castro G.t., Blance S.E. and Ferretti F.H., Internet
electro. J. Mol.Des.,3(11): 684-703 (2004).
28. Takahama U., Koga Y., Hirota S. and
Yamauchi R., Food Chemistry, 126(4): 1808–1811
(2011). (IVSL).
30. Lin C.M., Biochem Biophys Res Commun.,
294(1): 67-72 (2002).
31. Hawkes TR., George GN. and Bray RC.,
Biochem. J., 218(3): 961-968 (1984).
32. Skibo EB., Biochemistry, 25(15): 4189-4194
(1986).
33. Cengiz S., Cavas L., Yurdakoc K. and Aksu S.,
Turk. J. Biochem., 37 (4):445–45 (2012).
34. Nile S.H. and Khobragade CN., Indian Journal of
Natural Products and Resources, 2(1):52-58 (2011).
35. Barnes P.J., N.Engl.J.Med., 343:269-280(2000).
36. Seis H., Ann. NY. Acad. Sci., 669: 7-20 (1992).