Inferring mitochondrial and cytosolic metabolism by coupling isotope tracing and deconvolution.
Nat Commun
; 14(1): 7525, 2023 11 18.
Article
en En
| MEDLINE
| ID: mdl-37980339
ABSTRACT
The inability to inspect metabolic activities within distinct subcellular compartments has been a major barrier to our understanding of eukaryotic cell metabolism. Previous work addressed this challenge by analyzing metabolism in isolated organelles, which grossly bias metabolic activity. Here, we describe a method for inferring physiological metabolic fluxes and metabolite concentrations in mitochondria and cytosol based on isotope tracing experiments performed with intact cells. This is made possible by computational deconvolution of metabolite isotopic labeling patterns and concentrations into cytosolic and mitochondrial counterparts, coupled with metabolic and thermodynamic modelling. Our approach lowers the uncertainty regarding compartmentalized fluxes and concentrations by one and three orders of magnitude compared to existing modelling approaches, respectively. We derive a quantitative view of mitochondrial and cytosolic metabolic activities in central carbon metabolism across cultured cell lines without performing cell fractionation, finding major variability in compartmentalized malate-aspartate shuttle fluxes. We expect our approach for inferring metabolism at a subcellular resolution to be instrumental for a variety of studies of metabolic dysfunction in human disease and for bioengineering.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Respiración de la Célula
/
Mitocondrias
Límite:
Humans
Idioma:
En
Año:
2023
Tipo del documento:
Article