Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism
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 metho...
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| Format: | Article |
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Elsevier
2018
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| Online Access: | http://hdl.handle.net/1721.1/116785 https://orcid.org/0000-0002-7043-5013 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0002-1446-7256 |
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| author | Freinkman, Elizaveta Birsoy, Kıvanç Chen, Walter W. Wang, Tim Sabatini, David |
| author2 | Massachusetts Institute of Technology. Institute for Medical Engineering & Science |
| author_facet | Massachusetts Institute of Technology. Institute for Medical Engineering & Science Freinkman, Elizaveta Birsoy, Kıvanç Chen, Walter W. Wang, Tim Sabatini, David |
| author_sort | Freinkman, Elizaveta |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T11:47:10Z |
| format | Article |
| id | mit-1721.1/116785 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T11:47:10Z |
| publishDate | 2018 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1167852022-09-27T21:54:30Z Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism Freinkman, Elizaveta Birsoy, Kıvanç Chen, Walter W. Wang, Tim Sabatini, David Massachusetts Institute of Technology. Institute for Medical Engineering & Science Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Chen, Walter W. Wang, Tim Sabatini, David 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. 2018-07-05T13:50:12Z 2018-07-05T13:50:12Z 2016-08 2016-06 2018-07-03T18:30:52Z Article http://purl.org/eprint/type/JournalArticle 0092-8674 1097-4172 http://hdl.handle.net/1721.1/116785 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 https://orcid.org/0000-0002-7043-5013 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0002-1446-7256 http://dx.doi.org/10.1016/J.CELL.2016.07.040 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Freinkman, Elizaveta Birsoy, Kıvanç Chen, Walter W. Wang, Tim Sabatini, David Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism |
| title | Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism |
| title_full | Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism |
| title_fullStr | Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism |
| title_full_unstemmed | Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism |
| title_short | Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism |
| title_sort | absolute quantification of matrix metabolites reveals the dynamics of mitochondrial metabolism |
| url | http://hdl.handle.net/1721.1/116785 https://orcid.org/0000-0002-7043-5013 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0002-1446-7256 |
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