Carbon metabolism of Listeria monocytogenes growing inside macrophages
Open Access
- 9 July 2008
- journal article
- Published by Wiley in Molecular Microbiology
- Vol. 69 (4), 1008-1017
- https://doi.org/10.1111/j.1365-2958.2008.06337.x
Abstract
The intracellular metabolism of Listeria monocytogenes was studied by 13C‐isotopologue profiling using murine J774A.1 macrophages as host cells. Six hours after infection, bacteria were separated from the macrophages and hydrolyzed. Amino acids were converted into tert‐butyl‐dimethylsilyl derivatives and subjected to gas chromatography/mass spectrometry. When the macrophages were supplied with [U‐13C6]glucose prior to infection, but not during infection, label was detected only in Ala, Asp and Glu of the macrophage and bacterial protein with equal isotope distribution. When [U‐13C6]glucose was provided during the infection period, 13C label was found again in Ala, Asp and Glu from host and bacterial protein, but also in Ser, Gly, Thr and Val from the bacterial fraction. Mutants of L. monocytogenes defective in the uptake and catabolism of the C3‐metabolites, glycerol and/or dihydroxyacetone, showed reduced incorporation of [U‐13C6]glucose into bacterial amino acids under the same experimental settings. The 13C pattern suggests that (i) significant fractions (50–100%) of bacterial amino acids were provided by the host cell, (ii) a C3‐metabolite can serve as carbon source for L. monocytogenes under intracellular conditions and (iii) bacterial biosynthesis of Asp, Thr and Glu proceeds via oxaloacetate by carboxylation of pyruvate.Keywords
This publication has 29 references indexed in Scilit:
- Glycerol Metabolism and PrfA Activity inListeria monocytogenesJournal of Bacteriology, 2008
- Accurate Assessment of Amino Acid Mass Isotopomer Distributions for Metabolic Flux AnalysisAnalytical Chemistry, 2007
- Interference of Components of the Phosphoenolpyruvate Phosphotransferase System with the Central Virulence Gene Regulator PrfA ofListeria monocytogenesJournal of Bacteriology, 2007
- Carbon metabolism of intracellular bacteriaCellular Microbiology, 2006
- Some lessons fromRickettsiagenomicsFEMS Microbiology Reviews, 2005
- ListeriaPathogenesis and Molecular Virulence DeterminantsClinical Microbiology Reviews, 2001
- Elucidation of novel biosynthetic pathways and metabolite flux patterns by retrobiosynthetic NMR analysisFEMS Microbiology Reviews, 1998
- Biosynthetically Directed Fractional 13C-labeling of Proteinogenic Amino Acids. An Efficient Analytical Tool to Investigate Intermediary MetabolismEuropean Journal of Biochemistry, 1995
- Retrobiosynthetic analysis of carbon fixation in the phototrophic eubacterium Chloroflexus aurantiacusEuropean Journal of Biochemistry, 1993
- Mass isotopomer analysis: Theoretical and practical considerationsJournal of Mass Spectrometry, 1991