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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.

Original publication

DOI

10.1007/s00249-012-0844-2

Type

Journal article

Journal

Eur Biophys J

Publication Date

08/2012

Volume

41

Pages

675 - 679

Keywords

Animals, Cell-Free System, Fatty Acids, Hydrogen-Ion Concentration, Ion Channels, Ion Transport, Liposomes, Membrane Potentials, Mice, Mitochondrial Proteins, Protons, Purines, Uncoupling Protein 1