Journal of Bioenergetics and Biomembranes

, Volume 49, Issue 3, pp 273–279 | Cite as

A novel method for assessment of local pH in periplasmic space and of cell surface potential in yeast

  • Jaromír Plášek
  • David Babuka
  • Dana Gášková
  • Iva Jančíková
  • Jakub Zahumenský
  • Milan Hoefer
Article

Abstract

Yeast cells exhibit a negative surface potential due to negative charges at the cell membrane surface. Consequently, local concentrations of cations at the periplasmic membrane surface may be significantly increased compared to their bulk environment. However, in cell suspensions only bulk concentrations of cations can be measured directly. Here we present a novel method enabling the assessment of local pH at the periplasmic membrane surface which can be directly related to the underlying cell surface potential. In this proof of concept study using Saccharomyces cerevisiae cells with episomally expressed pH reporter, pHluorin, intracellular acidification induced by the addition of the protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP) was measured using synchronously scanned fluorescence spectroscopy (SSF). The analysis of titration curves revealed that the pH at the periplasmic surface of S. cerevisiae cells was about two units lower than the pH of bulk medium. This pH difference was significantly decreased by increasing the ionic strength of the bulk medium. The cell surface potential was estimated to amount to −130 mV. Comparable results were obtained also with another protonophore, pentachlorophenol (PCP).

Keywords

Yeast Cell surface potential Saccharomyces cerevisiae Cytosolic pH Periplasmic pH pHluorin 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Jaromír Plášek
    • 1
  • David Babuka
    • 1
  • Dana Gášková
    • 1
  • Iva Jančíková
    • 1
  • Jakub Zahumenský
    • 1
  • Milan Hoefer
    • 2
  1. 1.Faculty of Mathematics and PhysicsCharles UniversityPragueCzech Republic
  2. 2.Institute of Cellular and Molecular BotanyUniversity of BonnBonnGermany

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