Advertisement

An Ubiquinone-Binding Protein, A Constituent of Iron-Sulfur Fragment Isolated from Mitochondrial NADH-Ubiquinone Reductase (Complex I)

  • Hiroshi Suzuki
  • Takayuki Ozawa

Summary

An ubiquinone-binding protein (QP) was purified from mitochondrial NADH-ubiquinone reductase (Complex I). Complex I was separated into 3 fragments: a fraction of hydrophobic proteins, that of soluble ironsulfur protein (IP) and soluble NADH dehydrogenase of flavoprotein by a procedure involving the resolution with DOC and cholate, followed by ethanol and ammonium acetate fractionations. 41% of the total ubiquinone was recovered in the IP fragment which consisted of 12 polypeptides. The QP was purified from the IP fragment with a hydrophobic affinity chromatography. SDS-polyacrylamide gel electrophoresis and immunoblotting showed that the purified QP corresponded to 14-kDa polypeptide of the IP fragment and was a different protein from the QP (12.4kDa) in Complex III. The purified QP (14 kDa) contained one mol ubiquinone per mol. The ubiquinone-depleted IP fragment could rebind ubiquinone. These results indicate that an ubiquinone-binding site in Complex I is on the 14-kDa polypeptide of the IP fragment.

Keywords

Sodium Dodecyl Sulfate Ammonium Acetate Quinol Oxidase Beef Heart Mitochondrion Millimolar Extinction Coefficient 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Crane, L. (1977) Ann. Rev. Biochem. 46, 439–469CrossRefGoogle Scholar
  2. 2.
    Yu, C.A., Yu, L., and King, T.E. (1977) Biochem. Biophys. Res. Commun. 78, 259–265CrossRefGoogle Scholar
  3. 3.
    Wang, T., and King, T.E. (1982) Biochem. Biophys. Res. Commun. 104, 591–596CrossRefGoogle Scholar
  4. 4.
    Suzuki, H., and King, T. E. (1983) J. Biol. Chem. 258, 352–358Google Scholar
  5. 5.
    Suzuki, H., and Ozawa, T. (1984) Biochem. Intl. 9, 563–568Google Scholar
  6. 6.
    Suzuki, H., and Ozawa, T. (1984) Biochem. Biophys. Res. Commun. 124, 889–895CrossRefGoogle Scholar
  7. 7.
    Hatefi, Y., Haavik, A.G., and Griffiths, D.E. (1962)J. Biol. Chem. 231 1676–1680Google Scholar
  8. 8.
    Redfearn, E.R. (1967) Methods Enzymol. 10, 381–384CrossRefGoogle Scholar
  9. 9.
    Merle, P., and Kadenbach, B. (1980) Eur. J. Biochem. 105, 499–507CrossRefGoogle Scholar
  10. 10.
    Tamka, M., Nishikimi, M., Suzuki, H., Ozawa, T., Okino, E., and Takahashi, H. (1986) Biochem. Biophys. Res. Commun. in pressGoogle Scholar
  11. 11.
    Sakurai, T., Shimomura, Y., Nishikimi, M., and Ozawa, T. (1986) Biochem. Biophys. Res. Commun. 136, 376–380CrossRefGoogle Scholar
  12. 12.
    Towbin, H., Staehelin, T., and Gordon, J. (1979) Proc. Natl. Acad. Sei. USA 26, 4350–4354CrossRefGoogle Scholar
  13. 13.
    Galante, Y.M., and Hatefi, Y. (1978) Methods Enzymol. 53, 15–21CrossRefGoogle Scholar
  14. 14.
    Ragan, C.I., Galante, Y.M., and Hatefi, Y. (1982) Biochemistry, 21 2518–2524CrossRefGoogle Scholar
  15. 15.
    Shimomura, Y., Nishikimi, M., and Ozawa, T. (1986) Anal. Biochem. 153, 126–131CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Hiroshi Suzuki
    • 1
  • Takayuki Ozawa
    • 1
  1. 1.Department of Biomedical Chemistry Faculty of MedicineUniversity of NagoyaNagoya 466Japan

Personalised recommendations