Tracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells.

TitleTracing Compartmentalized NADPH Metabolism in the Cytosol and Mitochondria of Mammalian Cells.
Publication TypeJournal Article
Year of Publication2014
AuthorsLewis CA, Parker SJ, Fiske BP, McCloskey D, Gui DY, Green CR, Vokes NI, Feist AM, Heiden MGVander, Metallo CM
JournalMol Cell
PubMed Date2014 May 28
ISSN1097-4164
Abstract

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.

Alternate JournalMol. Cell
PubMed ID24882210
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