Combined In-Cell NMR and Simulation Approach to Probe Redox-Dependent Pathway Control
| Autor: | Marta R. A. Matos, Pernille Rose Jensen, Nikolaus Sonnenschein, Sebastian Meier |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Cellular metabolism
Chemistry 010401 analytical chemistry Nuclear magnetic resonance spectroscopy Metabolism Saccharomyces cerevisiae Carbon-13 NMR Molecular Dynamics Simulation 010402 general chemistry NAD 01 natural sciences Redox 0104 chemical sciences Analytical Chemistry Kinetics Biophysics Glycolysis NAD+ kinase Nuclear Magnetic Resonance Biomolecular Oxidation-Reduction Intracellular Cells Cultured Hexoses |
| Zdroj: | Analytical chemistry. 91(8) |
| ISSN: | 1520-6882 |
| Popis: | Dynamic response of intracellular reaction cascades to changing environments is a hallmark of living systems. As metabolism is complex, mechanistic models have gained popularity for describing the dynamic response of cellular metabolism and for identifying target genes for engineering. At the same time, the detailed tracking of transient metabolism in living cells on the subminute time scale has become amenable using dynamic nuclear polarization-enhanced 13C NMR. Here, we suggest an approach combining in-cell NMR spectroscopy with perturbation experiments and modeling to obtain evidence that the bottlenecks of yeast glycolysis depend on intracellular redox state. In pre-steady-state glycolysis, pathway bottlenecks shift from downstream to upstream reactions within a few seconds, consistent with a rapid decline in the NAD+/NADH ratio. Simulations using mechanistic models reproduce the experimentally observed response and help identify unforeseen biochemical events. Remaining inaccuracies in the computation... |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|