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Question: in the figure from the article by thompson and deberardinis...

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In the Figure from the article by Thompson and DeBerardinis (shown below), it is proposed to feed stable isotopically labeled (Bis-1,6-13C) glucose and then to determine the amount of L-lactate and Actyl-CoA with 13C. The amount of 13C metabolites can be determined on the basis of their lighter weight using mass spec and related to normal or cancer states.

i) Trace the isotopically labeled carbon by pathway (only describe starting material and product, not every intermediate is required.) to the two metabolites analyzed (lactate, acetyl CoA).

ii) Explain why those 2-carbons have to be labeled.

iii) If more labeled glucose goes to L-lactate than to acetyl CoA, identify the pathways that are activated and why more L-lacte than acetyl CoA is observed.


iv) What would an increased ratio of labeled L-lactate to acetyl CoA imply for: glucose consumption and local tissue pH?

Figure Metabolic Flux Analysis: (Yes, the structures for pyruvate and lactate are incorrect on the left; they are drawn correctly on the right. HO OH HO он Introduce label into subject (oral, intravenous) но Pyruvate O- Pyruvate PDH Other nutrients Metabolism COA SCOA он Lactate >90% labeled Acetyl-COA <25% labeled OH L-Lactate Acetyl CoA Isolate informative metabolites from tissue of interest Analyze for abundance and 13C enrichment Quantify metabolic flux

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