A Novel Method for Assessment of Local PH in Periplasmic Space and of Cell Surface Potential in Yeast

Overview

Journal J Bioenerg Biomembr

Publisher Springer

Specialties Biochemistry
Biology
Endocrinology

Date 2017 Apr 14

PMID 28405872

Citations 1

Authors

Jaromir Plasek

David Babuka

Dana Gaskova

Iva Jancikova

Jakub Zahumensky

Milan Hoefer

Affiliations

Soon will be listed here.

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).

Citing Articles

H translocation by weak acid uncouplers is independent of H electrochemical gradient.

Plasek J, Babuka D, Hoefer M J Bioenerg Biomembr. 2017; 49(5):391-397.

PMID: 28900787 DOI: 10.1007/s10863-017-9724-x.

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