Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism
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nihms810319.pdf
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Author(s)
Freinkman, Elizaveta
Birsoy, Kıvanç
Chen, Walter W.
Wang, Tim
Sabatini, David
Date Issued
August 2016
Journal
Cell
Publisher
Elsevier
Citation
Chen, Walter W. et al. “Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism.” Cell 166, 5 (August 2016): 1324–1337 © 2016 Elsevier Inc
Version
Author's final manuscript
Abstract
Mitochondria house metabolic pathways that impact most aspects of cellular physiology. While metabolite profiling by mass spectrometry is widely applied at the whole-cell level, it is not routinely possible to measure the concentrations of small molecules in mammalian organelles. We describe a method for the rapid and specific isolation of mitochondria and use it in tandem with a database of predicted mitochondrial metabolites (“MITObolome”) to measure the matrix concentrations of more than 100 metabolites across various states of respiratory chain (RC) function. Disruption of the RC reveals extensive compartmentalization of mitochondrial metabolism and signatures unique to the inhibition of each RC complex. Pyruvate enables the proliferation of RC-deficient cells but has surprisingly limited effects on matrix contents. Interestingly, despite failing to restore matrix NADH/NAD balance, pyruvate does increase aspartate, likely through the exchange of matrix glutamate for cytosolic aspartate. We demonstrate the value of mitochondrial metabolite profiling and describe a strategy applicable to other organelles.
MIT Department
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/J.CELL.2016.07.040
