Journal of Bioenergetics and Biomembranes

, Volume 42, Issue 5, pp 387–393

Electrogenic plasma membrane H+-ATPase activity using voltage sensitive dyes

  • Steve Amoroso
  • Ronald J. Clarke
  • Anthony Larkum
  • Rosanne Quinnell
Article

DOI: 10.1007/s10863-010-9306-7

Cite this article as:
Amoroso, S., Clarke, R.J., Larkum, A. et al. J Bioenerg Biomembr (2010) 42: 387. doi:10.1007/s10863-010-9306-7
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Abstract

Fast responding voltage sensitive dyes, RH421 and di-4-ASPBS, were used to study the electrogenic properties of plant plasma membrane proton pumps on sealed plasma membrane vesicles extracted by two-phase partitioning from Beta vulgaris and Avena sativa cv Swan root material. Fluorescence spectroscopy in the presence of the dye RH421 (10.8 nM) was sufficiently sensitive to detect electrogenic activity of the extracted plant vesicles. The dye detection system could detect inhibition of electrogenic activity of vesicles by vanadate (75 μM) and stimulation by nigericin (0.5 μM). The newly developed dye di-4-ASPBS was less sensitive to detecting the electrogenic proton pump activity. This study represents an important innovation in plant biophysics as this class of fast responding voltage sensitive dyes have never to our knowledge been used to study electrogenic proton pump activity derived from plant membranes and represents a novel approach for carrying out such studies.

Keywords

Plant plasma membrane H+-ATPase Voltage sensitive dyes Two-phase partitioning 

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Steve Amoroso
    • 1
  • Ronald J. Clarke
    • 2
  • Anthony Larkum
    • 1
  • Rosanne Quinnell
    • 1
  1. 1.The School of Biological SciencesThe University of SydneySydneyAustralia
  2. 2.School of ChemistryThe University of SydneySydneyAustralia

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