title: Staphylococcus aureus alpha-toxin mediates general and cell type-specific changes in metabolite concentrations of immortalized human airway epithelial cells.
abstract: Staphylococcus aureus alpha-toxin (Hla) is a potent pore-forming cytotoxin that plays an important role in the pathogenesis of S. aureus infections, including pneumonia. The impact of Hla on the dynamics of the metabolome in eukaryotic host cells has not been investigated comprehensively. Using 1H-NMR, GC-MS and HPLC-MS, we quantified the concentrations of 51 intracellular metabolites and assessed alterations in the amount of 25 extracellular metabolites in the two human bronchial epithelial cell lines S9 and 16HBE14o- under standard culture conditions and after treatment with sub-lethal amounts (2 µg/ml) of recombinant Hla (rHla) in a time-dependent manner. Treatment of cells with rHla caused substantial decreases in the concentrations of intracellular metabolites from different metabolic pathways in both cell lines, including ATP and amino acids. Concomitant increases in the extracellular concentrations were detected for various intracellular compounds, including nucleotides, glutathione disulfide and NAD+. Our results indicate that rHla has a major impact on the metabolome of eukaryotic cells as a consequence of direct rHla-mediated alterations in plasma membrane permeability or indirect effects mediated by cellular signalling. However, cell-specific changes also were observed. Glucose consumption and lactate production rates suggest that the glycolytic activity of S9 cells, but not of 16HBE14o- cells, is increased in response to rHla. This could contribute to the observed higher level of resistance of S9 cells against rHla-induced membrane damage.
authors.affiliation: Institute of Biochemistry, University of Greifswald, Greifswald, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Gierok
authors.affiliation: Competence Center Functional Genomics, Junior Research Group Pathoproteomics, University of Greifswald, Greifswald, Germany.
authors.forename: Manuela
authors.initials: M
authors.lastname: Harms
authors.affiliation: Competence Center Functional Genomics, Junior Research Group Pathoproteomics, University of Greifswald, Greifswald, Germany.
authors.forename: Erik
authors.initials: E
authors.lastname: Richter
authors.affiliation: Animal Physiology and Biochemistry, Zoological Institute and Museum, University of Greifswald, Greifswald, Germany.
authors.forename: Jan-Peter
authors.initials: JP
authors.lastname: Hildebrandt
authors.affiliation: Institute of Biochemistry, University of Greifswald, Greifswald, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Lalk
authors.affiliation: Competence Center Functional Genomics, Junior Research Group Pathoproteomics, University of Greifswald, Greifswald, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Mostertz
authors.affiliation: Competence Center Functional Genomics, Junior Research Group Pathoproteomics, University of Greifswald, Greifswald, Germany.
authors.forename: Falko
authors.initials: F
authors.lastname: Hochgräfe
mesh_terms: D001427:Bacterial Toxins / Q000633:toxicity*; D001980:Bronchi / Q000166:cytology*; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects; D002952:Citric Acid Cycle / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D006019:Glycolysis / Q000187:drug effects; D006460:Hemolysin Proteins / Q000633:toxicity*; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D055432:Metabolomics; D009711:Nucleotides / Q000378:metabolism; D025341:Principal Component Analysis; D011994:Recombinant Proteins / Q000633:toxicity
chemical_list: D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D007773:Lactates; D009711:Nucleotides; D011994:Recombinant Proteins; C021091:staphylococcal alpha-toxin; D005973:Glutamine; D005947:Glucose
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