The Journal of Membrane Biology

, Volume 96, Issue 2, pp 165–173 | Cite as

Procedure using voltage-sensitive spin-labels to monitor dipole potential changes in phospholipid vesicles: The estimation of phloretin-induced conductance changes in vesicles

  • Walter R. Perkins
  • David S. Cafiso
Articles

Summary

Voltage-sensitive membrane potential probes were used to monitor currents resulting from positive or negative charge movement across small and large unilamellar phosphatidylcholine (PC) vesicles. Positive currents were measured for the paramagnetic phosphonium ion or for K+-valinomycin. Negative currents were indirectly measured for the anionic proton carriers CCCP and DNP by monitoring transmembrane proton currents. Phloretin, a compound that is believed to decrease dipole fields in planar bilayers, increases positive currents and decreases negative currents when added to egg PC vesicles. In these vesicles, positive currents are increased by phloretin addition to a much larger degree than CCCP currents are reduced. This asymmetry, with respect to the sign of the charge carrier, is apparently not the result of changes in the membrane dielectric constant. It is most easily explained by deeper binding minima at the membrane-solution interface for the CCCP anion, when compared to the phosphonium. The measured asymmetry and the magnitudes of the current changes are consistent with the predictions of a point dipole model. The use of potential-sensitive probes to estimate positive and negative currents, provides a methodology to monitor changes in the membrane dipole potential in vesicle systems.

Key Words

dipole potential phospholipid vesicles spin labels CCCP membrane potentials phloretin 

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Copyright information

© Springer-Verlag New York Inc. 1987

Authors and Affiliations

  • Walter R. Perkins
    • 1
  • David S. Cafiso
    • 1
  1. 1.Department of Chemistry and Biophysics ProgramUniversity of VirginiaCharlottesville

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