Assaying the proton transport and regulation of UCP1 using solid supported membranes.

Abstract : The uncoupling protein 1 (UCP1) is a mitochondrial protein that carries protons across the inner mitochondrial membrane. It has an important role in non-shivering thermogenesis, and recent evidence suggests its role in human adult metabolism. Using rapid solution exchange on solid supported membranes, we succeeded in measuring electrical currents generated by the transport activity of UCP1. The protein was purified from mouse brown adipose tissue, reconstituted in liposomes and absorbed on solid supported membranes. A fast pH jump activated the ion transport, and electrical signals could be recorded. The currents were characterized by a fast rise and a slow decay, were stable over time, inhibited by purine nucleotides and activated by fatty acids. This new assay permits direct observation of UCP1 activity in controlled cell-free conditions, and opens up new possibilities for UCP1 functional characterization and drug screening because of its robustness and its potential for automation.
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Article dans une revue
Biophysics of structure and mechanism, Springer-Verlag, 2012, 41 (8), pp.675-9
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http://hal.univ-grenoble-alpes.fr/hal-01179678
Contributeur : Frank Thomas <>
Soumis le : jeudi 23 juillet 2015 - 10:36:23
Dernière modification le : jeudi 15 mars 2018 - 15:46:06

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  • HAL Id : hal-01179678, version 1
  • PUBMED : 22847775

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Iulia Blesneac, Stéphanie Ravaud, Paul Machillot, Manuela Zoonens, Sandrine Masscheylen, et al.. Assaying the proton transport and regulation of UCP1 using solid supported membranes.. Biophysics of structure and mechanism, Springer-Verlag, 2012, 41 (8), pp.675-9. 〈hal-01179678〉

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