Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells
Eukaryotic cells compartmentalize biochemical processes in different organelles, often relying on metabolic cycles to shuttle reducing equivalents across intracellular membranes. NADPH serves as the electron carrier for the maintenance of redox homeostasis and reductive biosynthesis, with separate c...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2016
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| Online Access: | http://hdl.handle.net/1721.1/103913 https://orcid.org/0000-0002-6702-4192 https://orcid.org/0000-0003-0130-3428 https://orcid.org/0000-0002-6883-3805 |
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| author | Lewis, Caroline A. Parker, Seth J. Fiske, Brian Prescott McCloskey, Douglas Gui, Dan Yi Green, Courtney R. Feist, Adam M. Vander Heiden, Matthew G. Metallo, Christian M. Vokes, Natalie I. Lewis, Caroline |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Lewis, Caroline A. Parker, Seth J. Fiske, Brian Prescott McCloskey, Douglas Gui, Dan Yi Green, Courtney R. Feist, Adam M. Vander Heiden, Matthew G. Metallo, Christian M. Vokes, Natalie I. Lewis, Caroline |
| author_sort | Lewis, Caroline A. |
| collection | MIT |
| description | Eukaryotic cells compartmentalize biochemical processes in different organelles, often relying on metabolic cycles to shuttle reducing equivalents across intracellular membranes. NADPH serves as the electron carrier for the maintenance of redox homeostasis and reductive biosynthesis, with separate cytosolic and mitochondrial pools providing reducing power in each respective location. This cellular organization is critical for numerous functions but complicates analysis of metabolic pathways using available methods. Here we develop an approach to resolve NADP(H)-dependent pathways present within both the cytosol and the mitochondria. By tracing hydrogen in compartmentalized reactions that use NADPH as a cofactor, including the production of 2-hydroxyglutarate by mutant isocitrate dehydrogenase enzymes, we can observe metabolic pathway activity in these distinct cellular compartments. Using this system we determine the direction of serine/glycine interconversion within the mitochondria and cytosol, highlighting the ability of this approach to resolve compartmentalized reactions in intact cells. |
| first_indexed | 2024-09-23T09:37:38Z |
| format | Article |
| id | mit-1721.1/103913 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:37:38Z |
| publishDate | 2016 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1039132022-09-30T15:46:45Z Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells Lewis, Caroline A. Parker, Seth J. Fiske, Brian Prescott McCloskey, Douglas Gui, Dan Yi Green, Courtney R. Feist, Adam M. Vander Heiden, Matthew G. Metallo, Christian M. Vokes, Natalie I. Lewis, Caroline Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Lewis, Caroline A. Fiske, Brian Prescott Gui, Dan Yi Vokes, Natalie Vander Heiden, Matthew G. Eukaryotic cells compartmentalize biochemical processes in different organelles, often relying on metabolic cycles to shuttle reducing equivalents across intracellular membranes. NADPH serves as the electron carrier for the maintenance of redox homeostasis and reductive biosynthesis, with separate cytosolic and mitochondrial pools providing reducing power in each respective location. This cellular organization is critical for numerous functions but complicates analysis of metabolic pathways using available methods. Here we develop an approach to resolve NADP(H)-dependent pathways present within both the cytosol and the mitochondria. By tracing hydrogen in compartmentalized reactions that use NADPH as a cofactor, including the production of 2-hydroxyglutarate by mutant isocitrate dehydrogenase enzymes, we can observe metabolic pathway activity in these distinct cellular compartments. Using this system we determine the direction of serine/glycine interconversion within the mitochondria and cytosol, highlighting the ability of this approach to resolve compartmentalized reactions in intact cells. National Institutes of Health (U.S.) (NIH grant P30CA147882) National Institutes of Health (U.S.) (NIH grant U54- CA121852-09) National Institutes of Health (U.S.) (NIH grant R01CA168653) David H. Koch Institute for Integrative Cancer Research at MIT (Koch Institute/DFHCC Bridge Project) David H. Koch Institute for Integrative Cancer Research at MIT (Koch Institute Frontier Research) Burroughs Wellcome Fund Damon Runyon Cancer Research Foundation Kathy and Curt Marble Cancer Research Fund American Cancer Society (grant IRG #70-002) United States. Department of Defense (DOD grant W81XWH-13-1-0105) University of California, San Diego (University of California Cancer Research Coordinating Committee grant) Searle Scholars Program (Award) 2016-08-15T17:24:40Z 2016-08-15T17:24:40Z 2014-05 2014-04 Article http://purl.org/eprint/type/JournalArticle 10972765 http://hdl.handle.net/1721.1/103913 Lewis, Caroline A., Seth J. Parker, Brian P. Fiske, Douglas McCloskey, Dan Y. Gui, Courtney R. Green, Natalie I. Vokes, Adam M. Feist, Matthew G. Vander Heiden, and Christian M. Metallo. "Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells" Molecular Cell 55:2 (July 2014), pp. 253-263. https://orcid.org/0000-0002-6702-4192 https://orcid.org/0000-0003-0130-3428 https://orcid.org/0000-0002-6883-3805 en_US http://dx.doi.org/10.1016/j.molcel.2014.05.008 Molecular Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Lewis, Caroline A. Parker, Seth J. Fiske, Brian Prescott McCloskey, Douglas Gui, Dan Yi Green, Courtney R. Feist, Adam M. Vander Heiden, Matthew G. Metallo, Christian M. Vokes, Natalie I. Lewis, Caroline Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells |
| title | Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells |
| title_full | Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells |
| title_fullStr | Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells |
| title_full_unstemmed | Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells |
| title_short | Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells |
| title_sort | tracing compartmentalized nadph metabolism in the cytosol and mitochondria of mammalian cells |
| url | http://hdl.handle.net/1721.1/103913 https://orcid.org/0000-0002-6702-4192 https://orcid.org/0000-0003-0130-3428 https://orcid.org/0000-0002-6883-3805 |
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