Skip to main content
Log in

Estimation of Membrane Potential Δψ in Reconstituted Plasma Membrane Vesicles Using a Numerical Model of Oxonol VI Distribution

  • Published:
Journal of Bioenergetics and Biomembranes Aims and scope Submit manuscript

Abstract

A model of membrane potential-dependent distribution of oxonol VI to estimate the electrical potential difference Δψ across Schizosaccharomyces pombe plasma membrane vesicles (PMV) has been developed. Δψ was generated by the H+-ATPase reconstituted in the PMV. The model treatment was necessary since the usual calibration of the dye fluorescence changes by diffusion potentials (K+ + valinomycin) failed. The model allows for fitting of fluorescence changes at different vesicle and dye concentrations, yielding Δψ in ATP-energized PMV of 80 mV. The described model treatment to estimate Δψ may be applicable for other reconstituted membrane systems.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

REFERENCES

  • Apell, H.-J., and Bersch, B. (1987). “Oxonol VI as an optical indicator for membrane potentials in lipid vesicles,” Biochim. Biophys. Acta 903, 480–494.

    Google Scholar 

  • Bashford, C. L., and Smith, J. C. (1979). “The use of optical probes to monitor membrane potential,” Methods Enzymol. 55, 569–586.

    Google Scholar 

  • Bashford, C. L., Chance, B., and Smith, J. C. (1979). “The behavior of oxonol dyes in phospholipid dispersions,” Biophys. J. 25, 63–85.

    Google Scholar 

  • Beeler, T. J., Farmen, R. H., and Martonosi, A. N. (1981). “The mechanism of voltage-sensitive dye responses on sarcoplasmatic reticulum,” J. Membr. Biol. 62, 3–137.

    Google Scholar 

  • Cooper, C. E., Bruce, D., and Nicolls, P. (1990). “Use of oxonol V as a probe of membrane potential in proteoliposomes containing cytochrome oxidase in the submitochondrial orientation,” Biochem. 29, 3859–3865.

    Google Scholar 

  • Gläser, H.-U., and Höfer, M. (1987). “Ion-dependent generation of the electrochemical proton gradient ΔμH + in reconstituted plasma membrane vesicles from the yeast Metschnikowia reukaufii,” Biochim. Biophys. Acta 905, 287–294.

    Google Scholar 

  • Höfer, M., Mair, T., and Wernsdörfer, E. (1991). “Molecular biology and biotechnology,” In: Yeast (Prasad, R. ed.), Omega Scientific Publishers, New Delhi, pp. 239–253.

    Google Scholar 

  • Kletterer, B., Neumcke, B., and Läuger, P. (1971). “Transport mechanism of hydrophobic ions through lipid bilayer membranes,” J. Membr. Biol. 5, 225–245.

    Google Scholar 

  • Mair, T., and Höfer, M. (1988). “ATP-induced generation of pH-gradient and/or membrane potential in reconstituted plasma membrane vesicles from Schizosaccharomyces pombe,” Biochem. Int. 17, 593–604.

    Google Scholar 

  • Perlin, D. S., Kasamo, K., Brooker, R. J., and Slayman, C. W. (1984). “Electrogenic H+ translocation by the plasma membrane ATPase of Neurospora,” J. Biol. Chem. 259, 7884–7892.

    Google Scholar 

  • Plášek, J., and Sígler, K. (1995). “Slow fluorescent indicators of membrane potential: a survey of different approaches to probe response analysis,” J. Photochem. Photobiol. in press.

  • Pratap, P. R., Novak, T. S., and Freedman, J. C. (1990). “Two mechanisms by which fluorescent oxonols indicate membrane potential in human red blood cells,” Biophys. J. 57, 835–849.

    Google Scholar 

  • Scherman, D., and Henry, J. P. (1980). “Oxonol V as a probe of chromaffin granule membrane potentials,” Biochim. Biophys. Acta 599, 150–166.

    Google Scholar 

  • Smith, J. C. (1990). “Potential-sensitive molecular probes in membranes of bioenergetic relevance,” Biochim. Biophys. Acta 1016, 1–28.

    Google Scholar 

  • Smith, J. C., Hallidy, L., and Topp, M. R. (1981). “The behavior of the fluorescence lifetime and polarization of oxonol potential-sensitive extrinsic probes in solution and in beef heart submitochondrial particles,” J. Membr. Biol. 60, 173–185.

    Google Scholar 

  • Villalobo, A. (1990). “Reconstitution of ion-motive transport ATPases in artificial lipid membranes,” Biochim. Biophys. Acta 1017, 1–48.

    Google Scholar 

  • Wolk, U., and Höfer, M. (1987). “Interactions between cyanine dyes and yeast cells: do cyanine dyes act as membrane potential sensitive probes?” Biochem. Int. 14, 501–509.

    Google Scholar 

  • Zwolinsky, B. J., Eyring, H., and Reese, C. (1949). “Diffusion and membrane permeability,” J. Phys. Colloid Chem. 53, 1426–1453.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Portele, A., Lenz, J. & Höofer, M. Estimation of Membrane Potential Δψ in Reconstituted Plasma Membrane Vesicles Using a Numerical Model of Oxonol VI Distribution. J Bioenerg Biomembr 29, 603–609 (1997). https://doi.org/10.1023/A:1022439203461

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022439203461

Navigation