Absolute quantification of matrix metabolites reveals the dynamics of mitochondrial metabolism (2016)
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, which we use in tandem with a database of predicted mitochondrial metabolites (“MITObolome”) to measure the matrix concentrations of greater than 100 metabolites across various states of respiratory chain (RC) function. Disruption of the RC revealed extensive compartmentalization of mitochondrial metabolism and signatures unique to the inhibition of each RC complex. Pyruvate enables the proliferation of RC-deficient cells, but had surprisingly limited effects on matrix contents. Interestingly, despite failing to restore matrix NADH/NAD balance, pyruvate did 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.
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Publication Details
Authors
Walter W. Chen, Elizaveta Freinkman, Tim Wang, Kıvanç Birsoy, and David M. Sabatini
Journal
PubMed
https://pubmed.ncbi.nlm.nih.gov/27565352/
Full Text
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5030821/
Citation
Chen WW, Freinkman E, Wang T, Birsoy K, Sabatini DM. Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism. Cell. 2016 Aug 25;166(5):1324-1337.e11. doi: 10.1016/j.cell.2016.07.040. PMID: 27565352; PMCID: PMC5030821.