Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Standard

Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems. / Wielandt, Alex Green; Palmgren, Michael Broberg; Fuglsang, Anja Thoe; Günther-Pomorski, Thomas; Justesen, Bo Højen.

P-Type ATPases: methods and protocols. ed. / Maike Bublitz. Vol. Part II Springer, 2016. p. 171-180 (Methods in Molecular Biology, Vol. 1377).

Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review

Harvard

Wielandt, AG, Palmgren, MB, Fuglsang, AT, Günther-Pomorski, T & Justesen, BH 2016, Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems. in M Bublitz (ed.), P-Type ATPases: methods and protocols. vol. Part II, Springer, Methods in Molecular Biology, vol. 1377, pp. 171-180. https://doi.org/10.1007/978-1-4939-3179-8_17

APA

Wielandt, A. G., Palmgren, M. B., Fuglsang, A. T., Günther-Pomorski, T., & Justesen, B. H. (2016). Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems. In M. Bublitz (Ed.), P-Type ATPases: methods and protocols (Vol. Part II, pp. 171-180). Springer. Methods in Molecular Biology Vol. 1377 https://doi.org/10.1007/978-1-4939-3179-8_17

Vancouver

Wielandt AG, Palmgren MB, Fuglsang AT, Günther-Pomorski T, Justesen BH. Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems. In Bublitz M, editor, P-Type ATPases: methods and protocols. Vol. Part II. Springer. 2016. p. 171-180. (Methods in Molecular Biology, Vol. 1377). https://doi.org/10.1007/978-1-4939-3179-8_17

Author

Wielandt, Alex Green ; Palmgren, Michael Broberg ; Fuglsang, Anja Thoe ; Günther-Pomorski, Thomas ; Justesen, Bo Højen. / Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems. P-Type ATPases: methods and protocols. editor / Maike Bublitz. Vol. Part II Springer, 2016. pp. 171-180 (Methods in Molecular Biology, Vol. 1377).

Bibtex

@inbook{32d7a89892b74983807a601bdf423ecf,
title = "Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems",
abstract = "The activity of enzymes involved in active transport of matter across lipid bilayers can conveniently be assayed by measuring their consumption of energy, such as ATP hydrolysis, while it is more challenging to directly measure their transport activities as the transported substrate is not converted into a product and only moves a few nanometers in space. Here, we describe two methods for the measurement of active proton pumping across lipid bilayers and the concomitant formation of a membrane potential, applying the dyes 9-amino-6-chloro-2-methoxyacridine (ACMA) and oxonol VI. The methods are exemplified by assaying transport of the Arabidopsis thaliana plasma membrane H(+)-ATPase (proton pump), which after heterologous expression in Saccharomyces cerevisiae and subsequent purification has been reconstituted in proteoliposomes.",
keywords = "Adenosine Triphosphate, Aminoacridines, Arabidopsis, Biological Transport, Active, Gene Expression Regulation, Enzymologic, Hydrolysis, Isoxazoles, Lipid Bilayers, Membrane Potentials, Proton-Translocating ATPases, Saccharomyces cerevisiae, Journal Article, Research Support, Non-U.S. Gov't",
author = "Wielandt, {Alex Green} and Palmgren, {Michael Broberg} and Fuglsang, {Anja Thoe} and Thomas G{\"u}nther-Pomorski and Justesen, {Bo H{\o}jen}",
year = "2016",
doi = "10.1007/978-1-4939-3179-8_17",
language = "English",
isbn = "978-1-4939-3178-1",
volume = "Part II",
series = "Methods in Molecular Biology",
publisher = "Springer",
pages = "171--180",
editor = "Maike Bublitz",
booktitle = "P-Type ATPases",
address = "Switzerland",

}

RIS

TY - CHAP

T1 - Measuring H+ pumping and membrane potential formation in sealed membrane vesicle systems

AU - Wielandt, Alex Green

AU - Palmgren, Michael Broberg

AU - Fuglsang, Anja Thoe

AU - Günther-Pomorski, Thomas

AU - Justesen, Bo Højen

PY - 2016

Y1 - 2016

N2 - The activity of enzymes involved in active transport of matter across lipid bilayers can conveniently be assayed by measuring their consumption of energy, such as ATP hydrolysis, while it is more challenging to directly measure their transport activities as the transported substrate is not converted into a product and only moves a few nanometers in space. Here, we describe two methods for the measurement of active proton pumping across lipid bilayers and the concomitant formation of a membrane potential, applying the dyes 9-amino-6-chloro-2-methoxyacridine (ACMA) and oxonol VI. The methods are exemplified by assaying transport of the Arabidopsis thaliana plasma membrane H(+)-ATPase (proton pump), which after heterologous expression in Saccharomyces cerevisiae and subsequent purification has been reconstituted in proteoliposomes.

AB - The activity of enzymes involved in active transport of matter across lipid bilayers can conveniently be assayed by measuring their consumption of energy, such as ATP hydrolysis, while it is more challenging to directly measure their transport activities as the transported substrate is not converted into a product and only moves a few nanometers in space. Here, we describe two methods for the measurement of active proton pumping across lipid bilayers and the concomitant formation of a membrane potential, applying the dyes 9-amino-6-chloro-2-methoxyacridine (ACMA) and oxonol VI. The methods are exemplified by assaying transport of the Arabidopsis thaliana plasma membrane H(+)-ATPase (proton pump), which after heterologous expression in Saccharomyces cerevisiae and subsequent purification has been reconstituted in proteoliposomes.

KW - Adenosine Triphosphate

KW - Aminoacridines

KW - Arabidopsis

KW - Biological Transport, Active

KW - Gene Expression Regulation, Enzymologic

KW - Hydrolysis

KW - Isoxazoles

KW - Lipid Bilayers

KW - Membrane Potentials

KW - Proton-Translocating ATPases

KW - Saccharomyces cerevisiae

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1007/978-1-4939-3179-8_17

DO - 10.1007/978-1-4939-3179-8_17

M3 - Book chapter

C2 - 26695032

SN - 978-1-4939-3178-1

VL - Part II

T3 - Methods in Molecular Biology

SP - 171

EP - 180

BT - P-Type ATPases

A2 - Bublitz, Maike

PB - Springer

ER -

ID: 169733840