Biochemical Characterization of Boar Sperm Cytochrome Oxidase

  • Yau-Huei Wei
  • Yu-Huey Huang
  • Ching-Her Lin


Cytochrome oxidase was isolated from boar sperm midpieces by extraction and fractionation with ammonium sulfate in the presence of cholate. The enzyme was further purified to apparent homogeneity by the DEAE-cellulose column chromatography in the presence of 0.3% Triton X-100.

The purified enzyme exhibited oxidized and reduced optical spectra similar to those of the bovine heart and rat liver cytochrome oxidases. However, the sperm oxidase was found to contain much higher amounts of subunits I,II and III than the other smaller subunits. Interestingly, we found that the sperm oxidase was much more acid-stable than the bovine heart and rat liver counterparts. The optimum pH for the sperm oxidase catalyzing the electron transfer from ferrocytochrome c to cytochrome a was around pH 4.8, and those for the bovine heart and rat liver were 6.2 and 6.8 respectively. In addition, we found that Ca2+ ion (10-100 μM) inhibited the activity of the sperm oxidase, whereas stimulated those of the bovine heart and rat liver enzymes.

The peculiar properties of sperm cytochrome oxidase may be due to the fact that the well-packed chromosomes in the sperm headpiece do not function so that the nuclear gene-coded subunits are deficient in the sperm cytochrome oxidase. The finding that the sperm oxidase was more acid-stable is an example of structure-function coordination, and will be discussed from the viewpoint of chemiosmotic theory and the unique structure and functions of the sperm mitochondria.


Ammonium Sulfate Cytochrome Oxidase Intermembrane Space Bovine Heart Cytochrome Oxidase Activity 
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  1. 1.
    Capaldi, R. (1982) Biochim. Biophys. Acta 694, 291–306.Google Scholar
  2. 2.
    Azzi, A. (1980) Biochim. Biophys. Acta 594, 231–252.Google Scholar
  3. 3.
    Capaldi, R. A., Malatesta, F., and Darley-Usmar, V. M. (1983) Biochim. Biophys. Acta 726, 135–148.Google Scholar
  4. 4.
    Tzagoloff, A. (1982) Mitochondria,Academic Press, New York.Google Scholar
  5. 5.
    Hatefi, Y. (1985) Ann. Rev. Biochem. 54, 1015–1069.CrossRefGoogle Scholar
  6. 6.
    Ades, I.Z., and Cascarano, J. (1977) J. Bioenerg. Biomembr. 9,237- 253.CrossRefGoogle Scholar
  7. 7.
    Poy ton, R.O., and Schatz, G. (1975) J. Biol. Chem. 250, 752- 761.Google Scholar
  8. 8.
    Ludwig, B. (1980) Biochim. Biophys. Acta 594, 177–181.Google Scholar
  9. 9.
    Kadenbach, B., and Merle, P. (1981) FEBS Lett. 135, 1–11.CrossRefGoogle Scholar
  10. 10.
    Merle, P., and Kadenbach, B. (1980) Eur. J. Biochem. 105, 499- 507.CrossRefGoogle Scholar
  11. 11.
    Kadenbach, B., and Stroh, A. (1984) FEBS Lett. 173, 374–380.CrossRefGoogle Scholar
  12. 12.
    Goldberg, E., Aberna, D., Wheat, T.E., Urbanski, G.J., and Margoliash, E. (1977) Science 196, 1010–1012.CrossRefGoogle Scholar
  13. 13.
    Hennig, B. (1975) Eur. J. Biochem. 55, 167–183.CrossRefGoogle Scholar
  14. 14.
    Tombes, R.M., and Shapiro, B.M. (1985) Cell 41, 325–334.CrossRefGoogle Scholar
  15. 15.
    Hong, C.Y., Chiang, B.N., Ku, J., and Wei, Y.H. (1984) Lancet I,460–461.CrossRefGoogle Scholar
  16. 16.
    Hong, C.Y., Chiang, B.N’., Ku, J., Wei, Y.H., and Fong, J.C. (1985) Br. J. Clin. Pharmac. 19, 45–49.Google Scholar
  17. 17.
    Hong, C.Y., Chiang, B.N., Ku, J., Wei, Y.H., and Fong, J.C.(1985) Br. J. Clin. Pharmac. 19, 739–743.Google Scholar
  18. 18.
    Hancock, J.L., and Hovell, G.L.R. (1959) Vet. Res. 71, 664- 671.Google Scholar
  19. 19.
    Mohri, H, Mohri, T, and Ernster, L.(1965) Exp. Cell Res. 38, 217–246.CrossRefGoogle Scholar
  20. 20.
    Hecht, N.B., and Bradley, F.M. (1981) Gamete Res. 4, 433–449.CrossRefGoogle Scholar
  21. 21.
    Hartzell, C.R., Beinert, H., Van Gelder, B.F., and King, T.E. (1978) Meth. Enzymol. 53, 54–64.CrossRefGoogle Scholar
  22. 22.
    Smith, L., Davis, H.C., and Nava, M.E. (1979) Biochemistry 18, 3140–3146.CrossRefGoogle Scholar
  23. 23.
    Smith, L., and Conrad, H. (1956) Arch. Biochem. Biophys. 63, 403- 413.CrossRefGoogle Scholar
  24. 24.
    Yonetani, T. (1962) J. Biol. Chem. 237, 550–559.Google Scholar
  25. 25.
    Yu, C.A., Yu, L., and King, T.E. (1975)J. Biol. Chem. 250, 1383- 1392.Google Scholar
  26. 26.
    Swank, T.R., and Munkres, K.D. (1971) Anal. Biochem. 39, 462- 477.CrossRefGoogle Scholar
  27. 27.
    Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. (1951) J. Biol. Chem. 193, 265–275.Google Scholar
  28. 28.
    Hadek, R. (1969) Mammalian Fertilization, An Atlas of Ultrastructure,pp. 34–35, Academic Press, New York.Google Scholar
  29. 29.
    Hong, C.Y., Chiang, B.N., and Wei, Y. H. (1983) Br. J. Clin. Pharmac. 16, 487–490.Google Scholar
  30. 30.
    Lee, C.Y., Pegoraro, B., Topping, J.L., and Yuan, J.H. (1977) Mol. Cell Biochem. 18, 49–57.CrossRefGoogle Scholar
  31. 31.
    Winter, D.B., Bruyninckx, W.J., Foulke, F.G., Grinich, N.P., and Mason, H.S. (1980) J. Biol. Chem. 255, 11408–11414.Google Scholar
  32. 32.
    Hackernbrock, C.R. (1966) J. Cell Biol. 30, 269–297.CrossRefGoogle Scholar
  33. 33.
    Mitchell, P. (1966) Biol. Rev. 41, 445–502.CrossRefGoogle Scholar
  34. 34.
    Nicholls, D.G. (1978) Biochem. J. 176, 463–474.Google Scholar
  35. 35.
    Nicholls, D.G., and Akerman, K.E.O. (1982) Biochim. Biophys. Acta 683, 57–88.Google Scholar
  36. 36.
    Nedergaard, J. (1983) Eur. J. Biochem. 133, 185–191.CrossRefGoogle Scholar
  37. 37.
    Connolly, E., Nanberg, E., and Nedergaard, J. (1984) Eur. J. Biochem. 141, 187–193.CrossRefGoogle Scholar
  38. 38.
    Stroh, A., and Kadenbach, B. (1986) Eur. J. Biochem. 156, 199–204.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Yau-Huei Wei
    • 1
  • Yu-Huey Huang
    • 1
  • Ching-Her Lin
    • 1
  1. 1.Department of BiochemistryNational Yang-Ming Medical CollegeTaipeiTaiwan, R.O.C.

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