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

TitleAssessing the Mitochondrial Membrane Potential in Cells and In Vivo using Targeted Click Chemistry and Mass Spectrometry.
Publication TypeJournal Article
Year of Publication2015
AuthorsLogan, A, Pell, VR, Shaffer, KJ, Evans, C, Stanley, NJ, Robb, EL, Prime, TA, Chouchani, ET, Cochemé, HM, Fearnley, IM, Vidoni, S, James, AM, Porteous, CM, Partridge, L, Krieg, T, Smith, RAJ, Murphy, MP
JournalCell Metab
Date Published2015 Dec 16
ISSN1932-7420
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.

DOI10.1016/j.cmet.2015.11.014
Alternate JournalCell Metab.
Citation Key10.1016/j.cmet.2015.11.014
PubMed ID26712463
Grant List098565 / / Wellcome Trust / United Kingdom