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Assaying P-Type ATPases Reconstituted in Liposomes

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2016

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https://doi.org/10.1007/978-1-4939-3179-8_14
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BUBLITZ, Maike, ed.. P-Type ATPases : Methods and Protocols. New York, NY: Humana Press, 2016, pp. 127-156. Methods in Molecular Biology. 1377. ISBN 978-1-4939-3178-1. Available under: doi: 10.1007/978-1-4939-3179-8_14

Zusammenfassung

Reconstitution of P-type ATPases in unilamellar liposomes is a useful technique to study functional properties of these active ion transporters. Experiments with such liposomes provide an easy access to substrate-binding affinities of the ion pumps as well as to the lipid and temperature dependence of the pump current. Here, we describe two reconstitution methods by dialysis and the use of potential-sensitive fluorescence dyes to study transport properties of two P-type ATPases, the Na,K-ATPase from rabbit kidney and the K+-transporting KdpFABC complex from E. coli. Several techniques are introduced how the measured fluorescence signals may be analyzed to gain information on properties of the ion pumps.

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Fachgebiet (DDC)
570 Biowissenschaften, Biologie

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Reconstitution, Lipids, Detergents, Dialysis, BioBeads, Vesicles, Voltage-sensitive fluorescence dyes, Electrogenic ion transport, Pump current, Substrate dependence, Membrane

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ISO 690APELL, Hans-Jürgen, Bojana DAMNJANOVIC, 2016. Assaying P-Type ATPases Reconstituted in Liposomes. In: BUBLITZ, Maike, ed.. P-Type ATPases : Methods and Protocols. New York, NY: Humana Press, 2016, pp. 127-156. Methods in Molecular Biology. 1377. ISBN 978-1-4939-3178-1. Available under: doi: 10.1007/978-1-4939-3179-8_14
BibTex
@incollection{Apell2016Assay-33541,
  year={2016},
  doi={10.1007/978-1-4939-3179-8_14},
  title={Assaying P-Type ATPases Reconstituted in Liposomes},
  number={1377},
  isbn={978-1-4939-3178-1},
  publisher={Humana Press},
  address={New York, NY},
  series={Methods in Molecular Biology},
  booktitle={P-Type ATPases : Methods and Protocols},
  pages={127--156},
  editor={Bublitz, Maike},
  author={Apell, Hans-Jürgen and Damnjanovic, Bojana}
}
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