Purification of Functional F-ATP Synthase from Blue Native PAGE

Chiara Galber; Giulia Valente; Sophia von Stockum; Valentina Giorgio

doi:10.1007/978-1-4939-9018-4_20

Purification of Functional F-ATP Synthase from Blue Native PAGE

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

Authors

Chiara Galber¹, Giulia Valente¹, Sophia von Stockum^{2

3}, Valentina Giorgio⁴

Affiliations

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.

PMID: 30674031
DOI: 10.1007/978-1-4939-9018-4_20

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.

Publication types

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

MeSH terms

Animals
Blotting, Western / methods
Drosophila
Electrophoresis, Polyacrylamide Gel / methods
HeLa Cells
Humans
Mice
Mitochondria / enzymology*
Mitochondrial Proton-Translocating ATPases / chemistry
Mitochondrial Proton-Translocating ATPases / isolation & purification*
Mitochondrial Proton-Translocating ATPases / metabolism
Native Polyacrylamide Gel Electrophoresis / methods*
Protein Multimerization
Rosaniline Dyes / chemistry
Staining and Labeling / methods

Substances

Rosaniline Dyes
Coomassie blue
Mitochondrial Proton-Translocating ATPases