Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803

The FoF1 ATP synthase (ATPase) is one of the most important protein complexes in energy metabolism. The isolation of functional ATPase complexes is fundamental to address questions about its assembly, regulation, and functions. This protocol describes the purification of intact and active ATPase fro...

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Detalles Bibliográficos
Autores principales: Song, Kuo, Tholen, Stefan, Baumgartner, Desirée, Schilling, Oliver, Hess, Wolfgang R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9418591/
https://www.ncbi.nlm.nih.gov/pubmed/36039073
http://dx.doi.org/10.1016/j.xpro.2022.101623
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author Song, Kuo
Tholen, Stefan
Baumgartner, Desirée
Schilling, Oliver
Hess, Wolfgang R.
author_facet Song, Kuo
Tholen, Stefan
Baumgartner, Desirée
Schilling, Oliver
Hess, Wolfgang R.
author_sort Song, Kuo
collection PubMed
description The FoF1 ATP synthase (ATPase) is one of the most important protein complexes in energy metabolism. The isolation of functional ATPase complexes is fundamental to address questions about its assembly, regulation, and functions. This protocol describes the purification of intact and active ATPase from the model cyanobacterium Synechocystis sp. PCC 6803. Basis for purification is a 3×FLAG tag fused to the beta subunit. The ATPase is enzymatically active and its purity is demonstrated using mass spectrometry, denaturing, and blue-native PAGE. For complete details on the use and execution of this protocol, please refer to Song et al. (2022).
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spelling pubmed-94185912022-08-28 Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803 Song, Kuo Tholen, Stefan Baumgartner, Desirée Schilling, Oliver Hess, Wolfgang R. STAR Protoc Protocol The FoF1 ATP synthase (ATPase) is one of the most important protein complexes in energy metabolism. The isolation of functional ATPase complexes is fundamental to address questions about its assembly, regulation, and functions. This protocol describes the purification of intact and active ATPase from the model cyanobacterium Synechocystis sp. PCC 6803. Basis for purification is a 3×FLAG tag fused to the beta subunit. The ATPase is enzymatically active and its purity is demonstrated using mass spectrometry, denaturing, and blue-native PAGE. For complete details on the use and execution of this protocol, please refer to Song et al. (2022). Elsevier 2022-08-16 /pmc/articles/PMC9418591/ /pubmed/36039073 http://dx.doi.org/10.1016/j.xpro.2022.101623 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Song, Kuo
Tholen, Stefan
Baumgartner, Desirée
Schilling, Oliver
Hess, Wolfgang R.
Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803
title Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803
title_full Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803
title_fullStr Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803
title_full_unstemmed Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803
title_short Isolation of intact and active FoF1 ATP synthase using a FLAG-tagged subunit from the cyanobacterium Synechocystis sp. PCC 6803
title_sort isolation of intact and active fof1 atp synthase using a flag-tagged subunit from the cyanobacterium synechocystis sp. pcc 6803
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9418591/
https://www.ncbi.nlm.nih.gov/pubmed/36039073
http://dx.doi.org/10.1016/j.xpro.2022.101623
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