Topic "Metabolism in Cell Extracts"

Life can be broken down into a defined set of chemical reactions that are catalysed by enzymes and compose a large, dense, and highly connected network called metabolism. We propose the study of metabolism in active cell free extracts (CFE), obtained from the lysis of cells and subsequent separation of their soluble components. Nuclear Magnetic Resonance (NMR) is a well-adapted and non-destructive tool allowing the identification and quantification of small organic molecules in a complex matrix, both at a given time point or throughout chemical reactions in real time.

 
We have adapted NMR methods to monitor physicochemical parameters such as pH simultaneously to metabolite concentration during a chemical reaction, and have described NMR sequences for the specific identification of phosphorylated metabolites in complex biological samples. We study the glycolytic activity and pentose phosphate pathway/glycolysis partition in the CFE as a function of the cofactor pools. Our results confirm that cofactors can have a strong influence on glycolytic flux, echoing what has been observed in vivo, but also suggest further hypotheses on the role of cofactors on central metabolism.