Abstract
The lipophilic cation tetraphenylphosphonium (TPP+) is accumulated by human skin fibroblasts across both the plasma and mitochondrial membranes. We show here that TPP+ uptake is indeed greatly decreased under conditions leading to de-energization of mitochondria. The TPP+ accumulation in the presence of the proton ionophore FCCP has been used for determination of the plasma membrane potential across the plasma membrane, after correction for potential-independent binding of TPP+ to cellular components. Following this procedure, a value of 75 mV has been obtained. Through the amount of TPP+ released by FCCP treatment, an estimate of thein situ mitochondrial membrane potential has been made. Furthermore, we report that the mitochondrial component of TPP+ accumulation decreases with aging of fibroblast cultures.
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Abbreviations
- ΔΨm :
-
membrane potential across thein situ mitochondria
- ΔΨp :
-
membrane potential across the plasma membrane
- TPP+ :
-
tetraphenylphosphonium
- HEPES:
-
N-2-hydroxyethylpiperazineN′-2-ethanesulfonic acid
- FCCP:
-
carbonyl cyanidep-trifluoromethoxyphenylhydrazone
References
Åkerman, K. E. O. (1979).Biochim. Biophys. Acta 546, 341–347.
Arslan, P., Di Virgilio, F., Beltrame, M., Tsien, R. G., and Pozzan, T. (1985).J. Biol. Chem. 260, 2719–2727.
Bakeeva, L. E., Grinius, L. L., Jasaitis, A. A., Kuliene, V. V., Levitsky, O. D., Liberman, E. A., Severina, I. I., and Skulachev, V. P. (1970).Biochim. Biophys. Acta 216, 13–21.
Brown, M. S., and Goldstein, J. L. (1976).Science 191, 150–154.
Carruthers, A., and Melchior, D. L. (1986).Trend Biol. Sci. 11, 331–335.
Davis, R. J., Brand, M. D., and Martin, B. R. (1981).Biochem. J. 196, 133–147.
Eytan, G. D., Matheson, M. G., and Racker, E. (1976).J. Biol. Chem. 251, 6831–6837.
Fahn, S., Koval, G. J., and Albers, R. W. (1966).J. Biol. Chem. 241, 1882–1889.
Felber, S. M., and Brand, M. D. (1982).Biochem. J. 204, 577–585.
Fiskum, G. (1985).Cell Calcium 6, 25–37.
Gallo, R. I., Finkelstein, J. N., and Notter, R. H. (1984).Biochim. Biophys. Acta 771, 217–227.
Gazzola, G. C., Dall'Asta, V., Franchi-Gazzola, R., and White, H. F. (1981).Anal. Biochem. 115, 368–374.
Harsford, R. G. (1983).Biochim. Biophys. Acta 726, 41–80.
Hayflick, L., and Moorehead, P. S. (1961).Exp. Cell Res. 25, 585–597.
Hoek, J. B., Nicholls, D. G., and Williamson, J. R. (1980).J. Biol. Chem. 255, 1458–1464.
Kiefer, H., Blume, A. J., and Kaback, H. R. (1980).Proc. Natl. Acad. Sci. USA 77, 2200–2204.
Krämer, R., and Klingenberg, M. (1977).FEBS Lett. 82, 363–367.
Landini, M. P., and Rugolo, M. (1984).J. Gen. Virol. 65, 2269–2272.
Lichtshtein, D., Kaback, H. R., and Blume, A. J. (1979).Proc. Natl. Acad. Sci. USA 76, 650–654.
Lowry, O. M., Rosenbrough, N. H., Farr, A. L., and Randall, R. J. (1951).J. Biol. Chem. 193, 265–275.
Moolenaar, W. H., Harden, Y., de Laat, S. W., and Schlessinger, J. (1982).J. Biol. Chem. 257, 8502–8506.
Nicholls, D. G. (1974).Eur. J. Biochem. 50, 305–315.
Nicholls, D. G., Rugolo, M., Scott, I. G., and Meldolesi, J. (1982).Proc. Natl. Acad. Sci. USA 79, 7924–7928.
Neufeld, E. F., Lim, T. W., and Shapiro, L. J. (1975).Ann. Rev. Biochem. 44, 357–376.
Rodemann, H. P., and Bayreuther, K. (1986).Proc. Natl. Acad. Sci. USA 83, 2086–2090.
Rugolo, M., Dolly, O., and Nicholls, D. G. (1986a).Biochem. J. 233, 519–523.
Rugolo, M., Romeo, G., and Lenaz, G. (1986b).Biochem. Biophys. Res. Commun. 134, 233–239.
Russel, S. B., Russel, J. D., and Truppin, J. S. (1984).J. Biol. Chem. 259, 11464–11470.
Rydstrom, J., Kanner, N., and Racker, E. (1975).Biochem. Biophys. Res. Commun. 67, 831–839.
Scott, I. D., and Nicholls, D. G. (1980).Biochem. J. 186, 21–23.
Scott, I. D., Akerman, K. E. O., and Nicholls, D. G. (1980).Biochem. J. 192, 873–880.
Seegmiller, J. E. (1976).Adv. Hum. Genet. 6, 75–81.
Seemann, D., Furstenberger, G., and Marks, F. (1983).Eur. J. Biochem. 137, 485–494.
Singer, T. P., and Gutman, M. (1971).Adv. Enzymol. 34, 79–153.
Swift, M. R., and Todaro, G. L. (1968).J. Cell Physiol. 71, 61–63.
Vik, S. B., Georgevich, G., and Capaldi, R. A. (1981).Proc. Natl. Acad. Sci. USA 78, 1456–1460.
Villereal, M. L., and Cook, J. S. (1978).J. Biol. Chem. 253, 8257–8262.
Yechiel, E., Henis, Y. I., and Barenholz, Y. (1986).Biochim. Biophys. Acta 859, 95–104.
Zoccarato, F., Siliprandi, N., and Rugolo, M. (1983).Biochim. Biophys. Acta 734, 381–383.
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Rugolo, M., Lenaz, G. Monitoring of the mitochondrial and plasma membrane potentials in human fibroblasts by tetraphenylphosphonium ion distribution. J Bioenerg Biomembr 19, 705–718 (1987). https://doi.org/10.1007/BF00762304
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DOI: https://doi.org/10.1007/BF00762304