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Methods Mol Biol. 2019;1925:233-243. doi: 10.1007/978-1-4939-9018-4_20.

Purification of Functional F-ATP Synthase from Blue Native PAGE.

Author information

1
Neuroscience Institute and Department of Biomedical Sciences, CNR and University of Padua, Padua, Italy.
2
Department of Biology, University of Padua, Padua, Italy.
3
Fondazione Ospedale San Camillo, IRCCS, Venezia, Italy.
4
Neuroscience Institute and Department of Biomedical Sciences, CNR and University of Padua, Padua, Italy. vgiorgio@bio.unipd.it.

Abstract

In the presence of Ca2+, F-ATP synthase preparations eluted from Blue Native gels generate electrophysiological currents that are typical of an inner mitochondrial membrane mega-channel, the permeability transition pore. Here we describe an experimental protocol for purification of F-ATP synthase that allows to maintain the enzyme assembly and activity that are essential for catalysis and channel formation.

KEYWORDS:

Blue Native PAGE; F-ATP synthase; Gel purification; Lipid bilayer; Permeability transition

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