Abstract
Rat liver mitochondrial inner and outer membranes were subjected to the solubilizing effect of the nonionic detergent Triton X-100 under various conditions. After centrifugation, the supernatants (containing the solubilized fraction) and pellets were characterized chemically and/or ultrastructurally. The detergent seems to act by inducing a phase transition from membrane lamellae to mixed protein-lipid-detergent micelles. Different electron-micro-scopy patterns are shown by the inner membranes after treatment with different amounts of surfactant, whereas the corresponding images from outer membranes vary but slightly. Selective solubilization of various components is observed, especially in the case of the inner membrane. Some membrane lipids (e.g., cardiolipin) are totally solubilized at detergent concentrations when others, such as sphyngomyelin, remain in the membrane. Other inner-membrane components (flavins, cytochromes, coenzymeQ) show different solubilization patterns. This allows the selection of conditions for optimal solubilization of a given membrane component with some degree of selectivity. The influence of Triton X-100 on various mitochondrial inner-membrane enzyme activities was studied. The detergent seems to act especially through disruption of the topology of the functional complexes, although the activity of the individual enzymes appears to be preserved. Relatively simple enzyme activities, such as ATPase, are more or less solubilized according to the detergent concentration, whereas the more complex succinate-cytochromec reductase activity practically disappears even at low Triton X-100 concentrations.
Similar content being viewed by others
References
A. Helenius and K. Simons,Biochim. Biophys. Acta 415 (1975), 29.
R. Coleman, G. Holdsworth, and J. B. Finean,Biochim. Biophys. Acta 436 (1976), 38.
J. Yu, D.A. Fischman, and T.L. Stock,J. Supramol. Struct. 1 (1973), 233.
R. W. Egan, M. A. Jones, and A. L. Lehninger,J. Biol. Chem. 251 (1976), 4442.
P. W. Holloway,Anal. Biochem. 53 (1973), 304.
I. G. Gurtubay, E. Azagra, A. Gutierrez, J. C. G. Milicua, and F. M. Goñi,Biochem. Soc. Trans. 7 (1979), 72.
G. H. Hogeboom, inMethods in Enzymology S. P. Colowick and N. O. Kaplan, eds., Academic Press, New York, Vol. I (1955), p. 16.
D. F. Parsons, G. R. Williams, and B. Chance,Ann. N.Y. Acad. Sci. 137 (1966), 643.
O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall,J. Biol. Chem. 193 (1951), 265.
C. S. Wang and R. L. Smith,Anal. Biochem. 63 (1975), 414.
G. Fairbanks, T. L. Steck, and D. F. H. Wallach,Biochemistry 10 (1971), 2606.
B. Chance, inMethods in Enzymology S. P. Colowick and N. O. Kaplan, eds., Academic Press, New York, Vol IV (1957), p. 273.
J. N. Williams,Arch. Biochem. Biophys. 107 (1964), 537.
E. R. Redfearn, inMethods in Enzymology S. P. Colowick and N. O. Kaplan, eds., Academic Press, New York, Vol. X (1967), p. 381.
J. Rosing, D. A. Harris, E. C. Slater, and J. Kemp, Jr.,Biochim. Biophys. Acta 376 (1975), 13.
D. Zeigler and J. S. Rieske, inMethods in Enzymology S. P. Colowick and N. O. Kaplan, eds., Academic Press, New York, Vol. X (1967), p. 231.
H. D. Tisdale, inMethods in Enzymology S. P. Colowick and N. O. Kaplan, eds., Academic Press, New York, Vol. X (1967), p. 213
Y. Hatefi and J. S. Rieske, inMethods in Enzymology S. P. Colowick and N. O. Kaplan, eds., Academic Press, New York, Vol. X (1967), p. 225.
G. L. Sottocasa, B. Kuylenstierna, L. Ernster, and A. Bergstrand,J. Cell. Biol. 32 (1967), 415.
H. Weissbach, T. E. Smith, J. W. Daly, W. Bernhard, and S. J. Udenfriend,J. Biol. Chem. 235 (1960), 1160.
E. Santiago, S. J. Mule, C. M. Redman, M. R. Hokin, and L. E. Hokin,Biochim. Biophys. Acta 84 (1964), 550.
J. Folch, M. Lees, and G. H. Sloane-Stanley,J. Biol. Chem. 226 (1957), 497.
G. R. Bartlett,J. Biol. Chem. 234 (1959), 466.
N. M. Neskovic and D. M. Kostic,J. Chromatogr. 35 (1968), 297.
W. R. Morrison and L. M. Smith,J. Lipid Res. 5 (1964), 600.
P. J. Quinn,The Molecular Biology of Cell Membranes Macmillan, London (1976).
Y. Kagawa, inMethods in Membrane Biology E. D. Korn, ed., Plenum Press, New York, Vol. 1 (1974), p. 201.
W. W. Wainio, inThe Mammalian Mitochondrial Respiratory Chain B. Horecker, N. O. Kaplan, J. Marmur, and H. A. Scheraga, eds., Academic Press, New York (1970).
C. Tanford, inThe Hidrophobic Effect Wiley, New York (1973).
J. Steinhardt and J. A. Reynolds, inMultiple Equilibria in Proteins, New York (1969), p. 10.
R. Becker, A. Helenius, and K. Simons,Biochemistry 14 (1975), 1835.
W. O. Kwant and P. Seeman,Biochim. Biophys. Acta 183 (1969), 530.
C. Tanford,J. Mol. Biol. 69 (1972), 59.
J. J. Auborn, E. M. Eyring, and G. L. Chowles,Proc. Natl. Acad. Sci. U.S.A. 68 (1971), 1996.
A. Helenius and H. Soderlund,Biochim. Biophys. Acta 307 (1973), 287.
B. Loizaga, I. G. Gurtubay, J. M. Macarulla, F. M. Goñi, and J. C. Gomez,Biochem. Soc. Trans. 7 (1979), 70.
K. Inoue and T. Kitagawa,Biochim. Biophys. Acta. 426 (1976), 1.
F. H. Kirkpatrick, S. E. Gordesky, and G. V. Marinetti,Biochim. Biophys. Acta 345 (1974), 154.
M. Lepage,J. Lipid Res. 5 (1964), 587.
K. Simons, H. Garoff, A. Helenius, L. Kaariainen, and O. Renkonen, inPerspectives in Membrane Biology O. S. Estrada and C. Gitler, eds., Academic Press, New York (1974), p. 45.
I. A. Kozlov and V. P. Skulachev,Biochim. Biophys. Acta 463 (1977), 29.
E. Santiago, N. Lopez-Moratalla, and J. L. Segovia,Biochem. Biophys. Res. Comm. 53 (1973), 439.
L. Ernster, G. Sandri, T. Hundall, C. Carlsson, and K. Nordenbrand, inStructure and Function of Energy Transducing Membranes K. Van Dam and B. F. Van Gelder, eds., Elsevier, Amsterdam (1977), p. 209.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gurtubay, J.I.G., Goñi, F.M., Gómez-Fernández, J.C. et al. Triton X-100 solubilization of mitochondrial inner and outer membranes. J Bioenerg Biomembr 12, 47–70 (1980). https://doi.org/10.1007/BF00745012
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00745012