Advertisement

Science in China Series B: Chemistry

, Volume 41, Issue 6, pp 575–586 | Cite as

Microcalorimetric studies on the dismutation of superoxide anion catalyzed by superoxide dismutase

  • Yi Liang
  • Songsheng Qu
  • Cunxin Wang
  • Yuwen Liu
  • Zhiyong Wang
  • Zhaohua Song
  • Guolin Zou
Article

Abstract

Thermokinetics of the dismutation of superoxide anion (O 2 .- ) catalyzed by superoxide dismutase (SOD), in which the decomposition of hydrogen peroxide catalyzed by catalase is utilized as a source of oxygen and the autoxidation of pyrogallod as a source of the substrate (O 2 .- ), has been studied by microcalorimetry. The molar reaction enthalpies of the SOD reaction and the pyrogallol autoxidation have been measured as - 160.1 and - 218 kJ · mol-1, respectively, at 298. 15 K and pH 7.0. A novel method for the determination of SOD activity, the microcalorimetric assay for SOD, has been proposed. The experimental results showed that SOD had no effect on both kinetic parameters and the mechanism for the pyrogallol autoxidation. This autoxidation followed the second-order reaction kinetics in the presence of limited oxygen (the first order with respect to both pyrogallol and O2), and the second-order rate constants were hermined at 298.15 K and pH 8.0 to be 1.25 and 1.30 L·mol-1·s-1 in the absence and presence of SOD, respectively. A possible mechanism for the autoxidation of pyrogallol inhibited by SOD was also suggested.

Keywords

mlcrocalorimetry superoxide dismutase pyrogallol autoxidation thermoklnetics kinetics reaction mechanism 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    McCord, J. M., Fridovich, I., Superoxide dismutase: an enzymatic function for erythrocuprein (hemocuprein),J. Biol. Chem., 1969, 244(22): 6049.Google Scholar
  2. 2.
    Fang Yunzhong, Li Wenjie,Free Radicals and Enzymes (in Chinese), Beijing: Science Press, 1989.Google Scholar
  3. 3.
    Deroche, A., Morgenstern-Badarau, I., Cesario, M. et al., A seven-coordinate manganese (II) complex formed with a single tripodal heptadentate ligand as a new superoxide scavenger,J. Am. Chem. Soc., 1996, 118(19): 4567.CrossRefGoogle Scholar
  4. 4.
    Liu Weihua, Liang Yongtao, Luo Guimin et al., Preparation of the superoxide dismutase (SOD)-catalase conjugate and its properties,Chinese Chem. Bull. (in Chinese), 1994, (12): 36.Google Scholar
  5. 5.
    Banci, L., Carloni, P., Penna, G. L. et al., Molecular dynamics studies on superoxide dismutase and its mutants: the structural and functional role of Arg143,J. Am. Chem. Soc., 1992, 114(18): 6994.CrossRefGoogle Scholar
  6. 6.
    Uchida, K., Kawakishi, S., Identification of oxidized histidine generated at the active site of Cu, Zn-superoxide dismutase exposed to H2O2,J. Biol. Chem., 1994, 269(4): 2405.Google Scholar
  7. 7.
    Shu Zhangyong, Denaturation of copper zinc superoxide dismutase by guanidine hydrochloride,Acta. Biochim. Biophys. Sin. (in Chinese), 1996, 28(5): 499.Google Scholar
  8. 8.
    Wang Bangning, Tan Fu, Calorimetric study of thermal denaturation of superoxide dismutase,Science in China, Ser. B, 1994, 37(1): 6.Google Scholar
  9. 9.
    Marklund, S., Marklund, G., Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase,Eur. J. Biochem., 1974, 47(3): 469.CrossRefGoogle Scholar
  10. 10.
    Zou Guolin, Gui Xingfen, Zhong Xiaoling et al., Improvements in pyrogallol autoxidation method for the determination of SOD activity,Progress in Biochemistry and Biophysics (in Chinese), 1986, (4): 71.Google Scholar
  11. 11.
    Yuan Qinsheng, Wang Zhiyou, Weng Qingqing et al., Determination of superoxide dismutase activity by pyrogallol autoxidation method,China Pharmaceutical Industry (in Chinese), 1983. (1): 16.Google Scholar
  12. 12.
    Yuan Zhuobin, Gao Ruomei, Kinetics and mechanism of pyrogallol autoxidation,Chem. J. Chinese Universities (in Chinese), 1997, 18(9): 1438.Google Scholar
  13. 13.
    Chowdhry, B. Z., Beezer, A. E., Greenhow, E. J., Analysis of drugs by micrccalorimetry: isothermal power-conduction calorimetry and thermometric titrimetry,Talanta, 1983, 30(4): 209.CrossRefGoogle Scholar
  14. 14.
    Liang Yi, Wang Cunxin, Wu Dingquan et al., Microcalorimetry applied to the study of catalase reaction,Chem. J. Chinese Universities (in Chinese), 1995, 16(6): 924.Google Scholar
  15. 15.
    Liang Yi, Wang Cunxin, Wu Dingquan et al., Microcalorimetry applied to the study of product inhibition of single-substrate enzyme-catalyzed reactions,ActaChim. Sin. (in Chinese), 1996, 54(1): 38.Google Scholar
  16. 16.
    Liang Yi, Wang Cunxin, Wu Dingquan et al., Thermokinetic studies of the irreversible inhibition of single-substrate, enzymecatalyzed reactions,Thermochim. Acta, 1995, 268: 17.CrossRefGoogle Scholar
  17. 17.
    Liu Jingsong, Zeng Xiancheng, Deng Yu et al., Reduced extent method for thermokinetics (V)—Thermokinetics research method for consecutive first order reactions,Acta Chim. Sin. (in Chinese), 1994, 52: 663.Google Scholar
  18. 18.
    Gray, P., Chernistry of free radicals containing oxygen: part 2—Thermochemistry of the hydroxyl and hydroperoxyl radicals,Trans. Faraday Soc., 1959, 55: 408.CrossRefGoogle Scholar
  19. 19.
    Foner, S. N., Hudson, R. L., Mass spectrometry of the HO2 free radical,J. Chem. Phys., 1962, 36(100): 2681.CrossRefGoogle Scholar
  20. 20.
    Zou Guolin, Chen Dongming, Chen Lin et al., Studies on the linearization method for the quantitation of activity of supemxide dismutase,J. Wuhan University (Natural Science Edition) (in Chinese), 1997, 42(6): 779.Google Scholar

Copyright information

© Science in China Press 1998

Authors and Affiliations

  • Yi Liang
    • 1
  • Songsheng Qu
    • 1
  • Cunxin Wang
    • 1
  • Yuwen Liu
    • 1
  • Zhiyong Wang
    • 1
  • Zhaohua Song
    • 1
  • Guolin Zou
    • 2
  1. 1.Department of ChemistryWuhan UniversityWuhanChina
  2. 2.College of Life ScienceWuhan UniversityWuhanChina

Personalised recommendations