Manda karaciğer ve böbrek doku arginazının fotoinaktivasyonu ve kinetik özellikleri

Photoinactivation of buffalo liver and kidney tissue arginasses and its kinetic properties

Exposure of buffalo liver and kidney tissue arginases to a 150 w light source from a distance of 8-10 cm, in the presence of 0.05mM methylene blue, caused significant inhibition of arginase activity. It was determined that this inhibition occurred differently in liver and kidney tissues. Arginase activity was spectrophotometrically measured using the Thiosemicarbazide diacetyl-monoxime urea (TDMU) method. Following preincubation with MnCl2, exposure to a 150 w light source and the addition of methylene blue, the activity of arginase was found to be lower than that after preincubation with a 150 w light source and methylene blue and the addition of MnCl2. Addition of methylene blue to buffalo liver and kidney tissue did not reduce the activity of arginase when the enzymatic reaction was carried out in darkness or in daylight, indicating that arginase is subject to photoinactivation by methylene blue. Enzyme inhibition was reduced 45-48% to 63% in the liver tissue and 30-40% in the kidney tissue. The inhibitory effect of methylene blue on buffalo liver and kidney tissue of arginase and its kinetic properties were studied. The inhibition was found to be noncompetitive. Photoinactivation might show its effect by modifying imidazole groups of histidyl residues in arginase molecules.

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1. Fuentes J.M., Campo M.L. and Soler G.: Kinetics and Inhibition by Some Amino Acids of Lactating Rat Mammary Gland Arginase. Arch. Inter. Physiol. Bioch. Biophy. 1994, 102: 255-258.
2. Gargnata C.L. and Bond J.S.: Assay and Kinetic of Arginase. Analy. Biochem. 1986, 154: 388-394.
3. Ber E. and Muszynska G.: Chemical Modification of Rat Liver Arginase. Acta Biochemica Polonica. 1979, Vol 19: No: 1/2.
4. Nadolska-Lutyk J., Grabon W. and Porembska Z.: Arginase in Bull Testis. Acta Bioch. Polonica. 1990, 37: 377-384.
5. King J.: Practical Clinical Enzymology. D. Von Nostrand Company London. 220-225, 1965.
6. Bellin J.S. and Yankus A.: Influence of Dye Binding on the Sensitized Photooxidation of Amino Acids. Arch. Biochem. Biophys. 1968, 123: 18-28.
7. Konarska L., Tomaszewski and Rolczyk U.: Studies on L-Arginase in Developing Rat Small Intestine, Brain and Kidney. Biochem. Medic. Metab. Biol. 1986, 35: 170-178.
8. Geyer J.W. and Dabich D.: Rapid Method for Determination of Arginase Activity in Tissue Homogenate. Department of Biochemistry. 1971, 412-417.
9. Gornall A.G., Bardawill C.J. and David M.M.: Determination of Serum Proteins by Means of the Biüret Reaction. J. Biol. Chem:1975; 177: 751
10. Özdemir N. and Ozan S.: M. benedeni Arginaz›n›n Fotoinaktivasyonu ve Kinetik Özellikleri. Erciyes Üniv. Sa¤. Bil. Derg: 1993; 2(2): 152-157
11. Ozan S., Gürsu F. ve Gülen fi.: ‹nsan Tükürük Arginaz›n›n Fotoinaktivasyonu. Biyokimya Derg:1991; F.Ü. 16(3): 57-65
12. Ozan S. ve Gülen fi.: Farkl› Türlerin Organlar›nda Bulunan Arginazlar›n Metilen Mavisi ile Fotoinaktivasyonlar›. Do¤a-Tr. J. of Biology: 1991; 15 : 222-229.
13. Samuel M.: Sodium Nucleate Inhibition of Arginase Activity. Science: 1952, 115: 69-70
14. Ber E. and Muszynska G.: Chemical Modification of Rat Liver Arginase. Acta Biochem. Polonica: 1979; 26: 103-114.
15. Ray W.J. Jr: Photochemical Oxidation. In S.P. Colowick and N.O. Kaplan (Eds): Methods in Enzymology Academic Press New York. Vol. 11, pp. 490-497, 1967.