Fig. 3

Global mapping of 2HG and glutamine fate. Plots showing altered metabolomic features in colorectal carcinoma cells after introduction of either a labeled U-¹³C 2HG (n = 3 per group) or b U-¹³C glutamine (n = 3 per group) for 24 h. The data shown are from colorectal carcinoma cells harboring mutations in IDH1. The y axis shows the fold change of isotopic enrichment calculated by dividing the intensity of each feature in the labeled samples by the intensity of each feature in the non-labeled samples. Features found to be enriched are shown with enlarged circles for clarity. Features containing isotopic label, shown in red on the left, were determined to be adducts or fragments of 2HG. The inset shows the extracted ion chromatogram (i.e., the LC trace) of each feature related to 2HG, each having the same retention time and chromatographic shape as expected for adducts and fragments. In contrast to the 2HG experiment where no labeled products were detected, the label from glutamine was incorporated into hundreds of features. One of the labeled features is annotated as 2HG. The glutamine data serve as a positive control that our platform can successfully identify biotransformations of a labeled substrate