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The Effect of Buffers on Weak Acid Uptake by Vesicles
The assessment of weak acid membrane permeability (P(m)) frequently involves large unilamellar vesicles. It relies on measurements of the intravesicular pH drop, ΔpH(in), in response to a sudden augmentation of external acid concentration. However, ΔpH(in) may be primarily governed by non-instantane...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6406578/ https://www.ncbi.nlm.nih.gov/pubmed/30781892 http://dx.doi.org/10.3390/biom9020063 |
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author | Hannesschlaeger, Christof Barta, Thomas Pechova, Hana Pohl, Peter |
author_facet | Hannesschlaeger, Christof Barta, Thomas Pechova, Hana Pohl, Peter |
author_sort | Hannesschlaeger, Christof |
collection | PubMed |
description | The assessment of weak acid membrane permeability (P(m)) frequently involves large unilamellar vesicles. It relies on measurements of the intravesicular pH drop, ΔpH(in), in response to a sudden augmentation of external acid concentration. However, ΔpH(in) may be primarily governed by non-instantaneous protonation and deprotonation reactions of (i) the acid itself, (ii) the buffer molecules, and (iii) the fluorescent pH reporter dye. Moreover, buffer concentration and acid gradient also serve as determinants of ΔpH(in), as we show here. The uniexponential time constant (τ) of ΔpH(in)(t) is an invalid measure of P(m) as Arrhenius plots of P(m) and [Formula: see text] reveal different activation energies for acid influx. We calculate P(m) by fitting a mathematical model to experimental stopped-flow traces. The model takes into account not only the time course of total internal buffer capacity but also (i) water self-dissociation, (ii) volume changes due to acid induced osmotic water flow, and (iii) the spontaneous membrane proton leak. It allows extracting a P(m) of 30.8 ± 3.5 μm/s for formic acid for 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicles. |
format | Online Article Text |
id | pubmed-6406578 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-64065782019-03-13 The Effect of Buffers on Weak Acid Uptake by Vesicles Hannesschlaeger, Christof Barta, Thomas Pechova, Hana Pohl, Peter Biomolecules Article The assessment of weak acid membrane permeability (P(m)) frequently involves large unilamellar vesicles. It relies on measurements of the intravesicular pH drop, ΔpH(in), in response to a sudden augmentation of external acid concentration. However, ΔpH(in) may be primarily governed by non-instantaneous protonation and deprotonation reactions of (i) the acid itself, (ii) the buffer molecules, and (iii) the fluorescent pH reporter dye. Moreover, buffer concentration and acid gradient also serve as determinants of ΔpH(in), as we show here. The uniexponential time constant (τ) of ΔpH(in)(t) is an invalid measure of P(m) as Arrhenius plots of P(m) and [Formula: see text] reveal different activation energies for acid influx. We calculate P(m) by fitting a mathematical model to experimental stopped-flow traces. The model takes into account not only the time course of total internal buffer capacity but also (i) water self-dissociation, (ii) volume changes due to acid induced osmotic water flow, and (iii) the spontaneous membrane proton leak. It allows extracting a P(m) of 30.8 ± 3.5 μm/s for formic acid for 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) vesicles. MDPI 2019-02-13 /pmc/articles/PMC6406578/ /pubmed/30781892 http://dx.doi.org/10.3390/biom9020063 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Hannesschlaeger, Christof Barta, Thomas Pechova, Hana Pohl, Peter The Effect of Buffers on Weak Acid Uptake by Vesicles |
title | The Effect of Buffers on Weak Acid Uptake by Vesicles |
title_full | The Effect of Buffers on Weak Acid Uptake by Vesicles |
title_fullStr | The Effect of Buffers on Weak Acid Uptake by Vesicles |
title_full_unstemmed | The Effect of Buffers on Weak Acid Uptake by Vesicles |
title_short | The Effect of Buffers on Weak Acid Uptake by Vesicles |
title_sort | effect of buffers on weak acid uptake by vesicles |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6406578/ https://www.ncbi.nlm.nih.gov/pubmed/30781892 http://dx.doi.org/10.3390/biom9020063 |
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