Titlebook: Metabolic Reprogramming; Methods and Protocol Salvatore Papa,Concetta Bubici Book 2023 The Editor(s) (if applicable) and The Author(s), und

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Real-Time Monitoring of Hydrogen Peroxide Levels in Yeast and Mammalian Cells,and hypoxia response. H.O. generation and signaling are highly localized processes. Understanding the dynamics of this molecule inside intact cells with subcompartmental resolution is instrumental to unravel its role in cellular signaling. Different genetically encoded fluorescent sensors have been

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Tracer-Based Metabolic Analysis by NMR in Intact Perfused Human Liver Tissue,er disease (NAFLD) are available, but they fail to fully recreate the metabolic and cellular features of human disease. Thus, it is imperative to understand the metabolic interplay in human cells in the context of disease. We have applied nuclear magnetic resonance (NMR) spectroscopy approaches to e

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13C Isotope Labeling and Mass Spectrometric Isotope Enrichment Analysis in Acute Brain Slices,arious experimental conditions. This information is particularly relevant for the study and functional understanding of brain metabolic heterogeneity. We here describe a protocol for the analysis of metabolic enrichment analysis upon feeding of murine acute cerebellar slices with .C-labeled substrat

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Metabolite Analyses Using Nuclear Magnetic Resonance (NMR) Spectroscopy in Plasma of Patients with ht metabolites and lipids. NMR spectroscopy is a powerful approach when applied to the high-throughput analysis of plasma or serum samples allowing, in addition, the detection of total proteins, lipoproteins, and signals arising from the glycosylation of circulating acute-phase proteins. Here, we de

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Screening Kinase-Dependent Phosphorylation of Key Metabolic Reprogramming Regulators,thesis of new cell constituents needed for cell growth and division. The M2 isoform of pyruvate kinase (PKM2) catalyzes the last reaction of the glycolytic process. PKM2 promotes the transfer of a phosphate group from phosphoenolpyruvate (PEP) to ADP, generating ATP and releasing pyruvate. This rate

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Measuring the Oxidation State and Enzymatic Activity of Glyceraldehyde Phosphate Dehydrogenase (GAPsis. GAPDH catalyzes the nicotinamide adenine dinucleotide (NAD.)- and inorganic phosphate-dependent oxidation and phosphorylation of glyceraldehyde phosphate (GAP) to form 1,3-bisphosphoglycerate (BPG). As part of its mechanism of action, GAPDH employs a redox-sensitive cysteine that serves as a nu

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