Profiling of antioxidative enzyme expression induced by various food components using targeted proteome analysis.
A method was developed for targeted proteome analysis of the expression profile of a set of antioxidative enzymes in rat macrophages and applied to screen the antioxidative potential of several food components/foods.



Expression profiles of heme oxygenase 1, peroxiredoxin 1, thioredoxin reductase 1, glutathione reductase, glutathione-S transferase P1, and superoxide dismutase 1 were analyzed by nanoLC-MS/MS in selected scheduled reaction monitoring (sSRM) mode monitoring two to three peptides per protein and three transitions per peptide. Relative quantification was performed by metabolic labeling. The validated method was used to profile the activity of capsaicin, carnosol, diallyl trisulfide, maslinic acid, quercetin, sulforaphane, cinnamaldehyde and coffee extract to modulate the expression levels of antioxidative enzymes. Carnosol and sulforaphane most effectively induced protein expression, leading to upregulation of at least five out of the six antioxidative enzymes by a maximum factor of 22.80 ± 6.71 (heme oxygenase 1 by carnosol). Heme oxygenase 1 was most susceptible to nutritive modulation, whereas glutathione reductase expression rates were hardly affected.



Targeted mass proteome analysis allows comprehensive evaluation of antioxidative effects by food ingredients. Simultaneous expression analysis of a set of proteins provided valuable insights how various enzymes were differently affected by food components.
Food Chemistry Unit, Department of Chemistry and Pharmacy, Emil Fischer Center, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.
Sven
S
Bartenbacher
Food Chemistry Unit, Department of Chemistry and Pharmacy, Emil Fischer Center, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.
Christiane
C
Östreicher
Food Chemistry Unit, Department of Chemistry and Pharmacy, Emil Fischer Center, Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Erlangen, Germany.
Monika
M
Pischetsrieder
D045784:Abietanes / Q000494:pharmacology; D000818:Animals; D000975:Antioxidants / Q000032:analysis*; D002211:Capsaicin / Q000494:pharmacology; D002478:Cells, Cultured; D003069:Coffee; D005502:Food; D005980:Glutathione Reductase / Q000032:analysis; D051549:Glutathione S-Transferase pi / Q000032:analysis; D051547:Heme Oxygenase-1 / Q000032:analysis; D017879:Isothiocyanates / Q000494:pharmacology; D008264:Macrophages / Q000201:enzymology; D020543:Proteome; D051381:Rats; D013454:Sulfoxides; D013482:Superoxide Dismutase / Q000032:analysis; D054481:Thioredoxin Reductase 1 / Q000032:analysis
D045784:Abietanes; D000975:Antioxidants; D003069:Coffee; D017879:Isothiocyanates; D020543:Proteome; D013454:Sulfoxides; C068623:carnosol; D051547:Heme Oxygenase-1; D013482:Superoxide Dismutase; D005980:Glutathione Reductase; D054481:Thioredoxin Reductase 1; D051549:Glutathione S-Transferase pi; C496558:Gstp1 protein, rat; C016766:sulforaphane; D002211:Capsaicin
Antioxidative enzymes; Carnosol; Heme oxygenase 1; LC-MS/MS-SRM; Targeted mass spectrometry