Assessing the Mitochondrial Membrane Potential in Cells and In Vivo using Targeted Click Chemistry and Mass Spectrometry

Angela Logan; Victoria R Pell; Karl J Shaffer; Cameron Evans; Nathan J Stanley; Ellen L Robb; Tracy A Prime; Edward T Chouchani; Helena M Cochemé; Ian M Fearnley; Sara Vidoni; Andrew M James; Carolyn M Porteous; Linda Partridge; Thomas Krieg; Robin A J Smith; Michael P Murphy

doi:10.1016/j.cmet.2015.11.014

Assessing the Mitochondrial Membrane Potential in Cells and In Vivo using Targeted Click Chemistry and Mass Spectrometry

Cell Metab. 2016 Feb 9;23(2):379-85. doi: 10.1016/j.cmet.2015.11.014. Epub 2015 Dec 17.

Authors

Angela Logan¹, Victoria R Pell², Karl J Shaffer³, Cameron Evans³, Nathan J Stanley³, Ellen L Robb¹, Tracy A Prime¹, Edward T Chouchani⁴, Helena M Cochemé⁵, Ian M Fearnley¹, Sara Vidoni¹, Andrew M James¹, Carolyn M Porteous⁶, Linda Partridge⁷, Thomas Krieg², Robin A J Smith³, Michael P Murphy⁸

Affiliations

¹ Medical Research Council Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK.
² Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, UK.
³ Department of Chemistry, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand.
⁴ Medical Research Council Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK; Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, UK; Dana-Farber Cancer Institute and Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA; Department of Cell Biology, Harvard University Medical School, Boston, MA 02115, USA.
⁵ Institute of Healthy Ageing, Department of Genetics, Evolution, and Environment, University College London, Gower Street, London WC1E 6BT, UK; MRC Clinical Sciences Centre, Imperial College London, Du Cane Road, London W12 0NN, UK.
⁶ Department of Chemistry, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand; Department Department of Biochemistry, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand.
⁷ Institute of Healthy Ageing, Department of Genetics, Evolution, and Environment, University College London, Gower Street, London WC1E 6BT, UK.
⁸ Medical Research Council Mitochondrial Biology Unit, Hills Road, Cambridge CB2 0XY, UK. Electronic address: mpm@mrc-mbu.cam.ac.uk.

Abstract

The mitochondrial membrane potential (Δψm) is a major determinant and indicator of cell fate, but it is not possible to assess small changes in Δψm within cells or in vivo. To overcome this, we developed an approach that utilizes two mitochondria-targeted probes each containing a triphenylphosphonium (TPP) lipophilic cation that drives their accumulation in response to Δψm and the plasma membrane potential (Δψp). One probe contains an azido moiety and the other a cyclooctyne, which react together in a concentration-dependent manner by "click" chemistry to form MitoClick. As the mitochondrial accumulation of both probes depends exponentially on Δψm and Δψp, the rate of MitoClick formation is exquisitely sensitive to small changes in these potentials. MitoClick accumulation can then be quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This approach enables assessment of subtle changes in membrane potentials within cells and in the mouse heart in vivo.

Publication types

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

MeSH terms

Animals
Cell Line
Click Chemistry / methods*
Membrane Potential, Mitochondrial*
Mice, Inbred C57BL
Molecular Probes / metabolism
Tandem Mass Spectrometry / methods*

Substances

Molecular Probes

Abstract

Publication types

MeSH terms

Substances

Grants and funding