Bacterial metabolite labeling from U-13C mannitol.

Abstract

(A) Total fractional labeling enrichment (“TFLE” = 1 –unlabeled fraction) of different bacterial metabolites isolated from RAW264.7 cells at 12 hpi with MOI 100. Bars show averages of biological triplicates. CCM: central carbon metabolism, GSH: reduced glutathione; GSSG: oxidized glutathione. (B) Fractional 13C enrichment (long axis of bars, “Ratio”) of glycolytic bacterial metabolites isolated from the same experiment as (A) for the different isotopologues (M+0, M+1, M+2, …). U-13C mannitol (mtl) on the top right of the plot is the source for all labels in the isotopologues with higher masses (M+1, M+2, …). Dashed arrows depict lumped reactions. Note that glucose (glc) 6P and fructose (fru) 6P are indistinguishable by our analysis. Data are corrected for the natural 13C abundance and the 12C isotopic impurity of U-13C mtl and depict the average of biological triplicates; in (A) and (B) metabolites quantitatively affected by the enrichment protocol (S2E Fig) are denoted with *. The data underlying this figure can be found in S1 Data. DHAP, dihydroxyacetone phosphate; ED, Entner–Doudoroff; FBP, fructose 1,6 bisphosphate; G3P, glyceraldehyde 3P; hpi, hours post-infection; LPS, lipopolysaccharide; MOI, multiplicity of infection; mtl, mannitol; PEP, phosphoenolpyruvate; PP, pentose phosphate; TFLE, total fractional labeling enrichment.</p