The Journal of Membrane Biology

, Volume 7, Issue 1, pp 313–324 | Cite as

On the impermeability of the outer mitochondrial membrane to cytochromec

II. Studies on isolated membrane fragments
  • Lech Wojtczak
  • G. L. Sottocasa


Outer mitochondrial membranes isolated by the swelling-shrinkage sonication procedure of Sottocasaet al. [19, 20] forms small sealed vesicles. If cytochromec is present during the procedure it is trapped inside these vesicles and can not be washed out nor is accessible to external enzymes, e.g., cytochrome oxidase (EC or succinate-cytochromec reductase present as contamination by the inner membrane, but is fully accessible to rotenone-insensitive NADH-cytochromec reductase of the outer membrane. This indicates the impermeability of the outer mitochondrial membrane to cytochromec.

A modification of the original procedure for the separation of the outer mitochondrial membrane is described.


Human Physiology Mitochondrial Membrane Outer Membrane Cytochrome Oxidase Outer Mitochondrial Membrane 
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  1. 1.
    Allmann, D. W., Bachmann, E., Orme-Johnson, N., Tan, W. C., Green, D. E. 1968. Membrane systems of mitochondria. VI. Membranes of liver mitochondria.Arch. Biochem. Biophys. 125:981.PubMedGoogle Scholar
  2. 2.
    Galzigna, L., Sartorelli, L. 1968. Reduction of cytochromec by the mitochondrial respiratory chain across a semi-permeable membrane.Experientia 24:19.PubMedGoogle Scholar
  3. 3.
    Gornall, A. G., Bardawill, C. J., David, M. M. 1949. Determination of serum proteins by means of the biuret reaction.J. Biol. Chem. 177:751.Google Scholar
  4. 4.
    Hogeboom, G. H. 1955. Fractionation of cell components of animal tissues.In: Methods in Enzymology. S. P. Colowick and N. O. Kaplan, editors. Vol. 1, p. 16. Academic Press Inc., New York.Google Scholar
  5. 5.
    Jacobs, E. E., Sanadi, D. R. 1960. The reversible removal of cytochromec from mitochondria.J. Biol. Chem. 235:531.PubMedGoogle Scholar
  6. 6.
    Kimelberg, H. K., Lee, C.-P. 1969. Binding and electron transfer to cytochromec in artificial phospholipid membranes.Biochem. Biophys. Res. Commun. 34:784.PubMedGoogle Scholar
  7. 7.
    Klingenberg, M., Pfaff, E. 1966. Structural and functional compartmentation in mitochondria.In: Regulation of Metabolic Processes in Mitochondria. J. M. Tager, S. Papa, E. Quagliariello and E. C. Slater, editors. p. 180. Elsevier Publishing Co., Amsterdam.Google Scholar
  8. 8.
    Kuylenstierna, B., Nicholls, D. G., Movmöller, S., Ernster, L. 1970. Effect of trypsin on mitochondrial and microsomal enzymes.Europ. J. Biochem. 12:419.PubMedGoogle Scholar
  9. 9.
    Lee, C.-P., Carlson, K. 1968. Binding of cytochromec to fragmented mitochondrial membranes.Fed. Proc. 27:828.Google Scholar
  10. 10.
    Lehninger, A. L. 1962. Water uptake and extrusion by mitochondria in relation to oxidative phosphorylation.Physiol. Rev. 42:467.PubMedGoogle Scholar
  11. 11.
    Lehninger, A. L. 1964. The Mitochondrion. W. A. Benjamin, New York-Amsterdam.Google Scholar
  12. 12.
    Malamed, S., Recknagel, R. O. 1959. The osmotic behaviour of the sucrose-inaccessible space of mitochondrial pellets from rat liver.J. Biol. Chem. 234:3027.PubMedGoogle Scholar
  13. 13.
    O'Brien, R. L., Brierley, G. 1965. Compartmentation of heart mitochondria. 1. Permeability characteristics of isolated beef heart mitochondria.J. Biol. Chem. 240:4527.PubMedGoogle Scholar
  14. 14.
    Parsons, D. F., Williams, G. R., Thompson, W., Wilson, D., Chance, B. 1967. Improvements in the procedure for purification of mitochondrial outer and inner membrane. Comparison of the outer membrane with smooth endoplasmic reticulum.In: Mitochondrial Structure and Compartmentation. E. Quagliariello, S. Papa, E. C. Slater and J. M. Tager, editors. p. 29. Adriatica Editrice, Bari.Google Scholar
  15. 15.
    Pfaff, E., Klingenberg, M., Ritt, E., Vogell, W. 1968. Korrelation des unspezifisch permeablen mitochondrialen Raumes mit dem “Intermembranraum”.Europ. J. Biochem. 5:222.PubMedGoogle Scholar
  16. 16.
    Schnaitman, C., Greenawalt, J. W. 1968. Enzymatic properties of inner and outer membranes of rat liver mitochondria.J. Cell Biol. 38:158.PubMedGoogle Scholar
  17. 17.
    Silva Lima, M., Nachbaur, G., Vignais, P. V. 1968. Localisation de la nucléoside diphosphate kinase, de l'adénylate kinase et de la GTP-AMP phosphotransférase dans des mitochondries de foie.Compt. Rend. Acad. Sci. (Paris), Ser. D 266:739.Google Scholar
  18. 18.
    Skulachev, V. P. 1969. Energy Accumulation in the Cell (in Russian). Nauka, Moscow.Google Scholar
  19. 19.
    Sottocasa, G. L., Kuylenstierna, B., Ernster, L., Bergstrand, A. 1967. An electron-transport system associated with the outer membrane of liver mitochondria.J. Cell. Biol. 32:415.PubMedGoogle Scholar
  20. 20.
    Sottocasa, G. L., Kuylenstierna, B., Ernster, L., Bergstrand, A. 1967. Separation and some enzymatic properties of the inner and outer membranes of rat liver mitochondria.In: Methods in Enzymology. R. W. Estabrook and M. E. Pullman, editors. Vol. 10, p. 448. Academic Press Inc., New York.Google Scholar
  21. 21.
    Sottocasa, G. L., Sandri, G. 1969. The permeability to pyridine nucleotides of microsomes and outer mitochondrial membranes from rat liver.In: Biochemistry of Intracellular Structures. Mitochondria and Endoplasmic Reticulum. L. Wojtczak, W. Drabikowski and H. Strzelecka-Golaszewska, editors. p. 7. Polish Scientific Publishers, Warszawa.Google Scholar
  22. 22.
    Tedeschi, H., Harris, D. L. 1955. The osmotic behaviour and permeability to non-electrolytes of mitochondria.Arch. Biochem. Biophys. 58:52.PubMedGoogle Scholar
  23. 23.
    Wojtczak, L., Zaluska, H. 1969. On the impermeability of the outer mitochondrial membrane to cytochromec. I. Studies on whole mitochondria.Biochim. Biophys. Acta 193:64.PubMedGoogle Scholar

Copyright information

© Springer-Verlag New York Inc. 1972

Authors and Affiliations

  • Lech Wojtczak
    • 1
  • G. L. Sottocasa
    • 1
  1. 1.Istituto di Chimica BiologicaUniversity of TriesteTriesteItaly

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