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

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

来源：Cell Metabolism, 2016, 23(2): 379-385.

DOI：10.1016/j.cmet.2015.11.014

The mitochondrial membrane potential (Delta Psi(m)) is a major determinant and indicator of cell fate, but it is not possible to assess small changes in Delta Psi(m) 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 Delta Psi(m) and the plasma membrane potential (Delta Psi(p)). 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 Delta Psi(m) and Delta Psi(p), 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.

• 出版日期2016-2-9