A novel method for assessment of local pH in periplasmic space and of cell surface potential in yeast
Language English Country United States Media print-electronic
Document type Journal Article
PubMed
28405872
DOI
10.1007/s10863-017-9710-3
PII: 10.1007/s10863-017-9710-3
Knihovny.cz E-resources
- Keywords
- Cell surface potential, Cytosolic pH, Periplasmic pH, Saccharomyces cerevisiae, Yeast, pHluorin,
- MeSH
- Spectrometry, Fluorescence methods MeSH
- Carbonyl Cyanide m-Chlorophenyl Hydrazone MeSH
- Hydrogen-Ion Concentration * MeSH
- Membrane Potentials * MeSH
- Methods MeSH
- Periplasm chemistry MeSH
- Saccharomyces cerevisiae chemistry cytology MeSH
- Green Fluorescent Proteins MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Carbonyl Cyanide m-Chlorophenyl Hydrazone MeSH
- PHluorin MeSH Browser
- Green Fluorescent Proteins MeSH
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).
Faculty of Mathematics and Physics Charles University Ke Karlovu 3 12116 Prague Czech Republic
Institute of Cellular and Molecular Botany University of Bonn Kirschallee 1 53115 Bonn Germany
See more in PubMed
Biochim Biophys Acta. 1979 Dec 6;548(3):579-95 PubMed
Cryo Letters. 2015 May-Jun;36(3):221-6 PubMed
Methods Enzymol. 1989;171:387-94 PubMed
Biotechnol Prog. 2002 Jul-Aug;18(4):831-8 PubMed
Nat Commun. 2014;5:3103 PubMed
Langmuir. 2014 Sep 2;30(34):10328-35 PubMed
Anal Chem. 2015 Oct 6;87(19):9600-4 PubMed
Plant Physiol. 2011 Feb;155(2):808-20 PubMed
ACS Appl Mater Interfaces. 2014 Sep 10;6(17):15329-34 PubMed
Chem Commun (Camb). 2016 Jun 14;52(50):7818-21 PubMed
Yeast. 2010 Jun;27(6):317-25 PubMed
J Membr Biol. 1971 Dec;4(1):227-51 PubMed
Biochim Biophys Acta. 2006 Aug;1757(8):913-30 PubMed
Physiol Rev. 1980 Jul;60(3):825-63 PubMed
Proc Natl Acad Sci U S A. 2016 Jul 19;113(29):8177-81 PubMed
Plant Physiol. 2008 Dec;148(4):2134-43 PubMed
Folia Microbiol (Praha). 1962 Mar;7:109-14 PubMed
Biophys J. 1986 Feb;49(2):459-68 PubMed
J Membr Biol. 1984;82(2):179-90 PubMed
Proc Natl Acad Sci U S A. 2014 Sep 2;111(35):12684-8 PubMed
Biochemistry. 1993 Sep 28;32(38):10057-66 PubMed
Methods Enzymol. 1986;127:678-96 PubMed
Nature. 1967 Nov 18;216(5116):718-9 PubMed
J Bacteriol. 2007 Aug;189(15):5601-7 PubMed
Biochim Biophys Acta. 1981 Dec;650(2-3):88-127 PubMed
Proc Natl Acad Sci U S A. 1968 Feb;59(2):484-90 PubMed
Springerplus. 2015 Nov 04;4:672 PubMed
Methods Enzymol. 1989;171:364-75 PubMed
Biochim Biophys Acta. 1979 Jul 10;547(1):91-102 PubMed
H+ translocation by weak acid uncouplers is independent of H+ electrochemical gradient
