title: Redox proteomics of the inflammatory secretome identifies a common set of redoxins and other glutathionylated proteins released in inflammation, influenza virus infection and oxidative stress.
abstract: Protein cysteines can form transient disulfides with glutathione (GSH), resulting in the production of glutathionylated proteins, and this process is regarded as a mechanism by which the redox state of the cell can regulate protein function. Most studies on redox regulation of immunity have focused on intracellular proteins. In this study we have used redox proteomics to identify those proteins released in glutathionylated form by macrophages stimulated with lipopolysaccharide (LPS) after pre-loading the cells with biotinylated GSH. Of the several proteins identified in the redox secretome, we have selected a number for validation. Proteomic analysis indicated that LPS stimulated the release of peroxiredoxin (PRDX) 1, PRDX2, vimentin (VIM), profilin1 (PFN1) and thioredoxin 1 (TXN1). For PRDX1 and TXN1, we were able to confirm that the released protein is glutathionylated. PRDX1, PRDX2 and TXN1 were also released by the human pulmonary epithelial cell line, A549, infected with influenza virus. The release of the proteins identified was inhibited by the anti-inflammatory glucocorticoid, dexamethasone (DEX), which also inhibited tumor necrosis factor (TNF)-α release, and by thiol antioxidants (N-butanoyl GSH derivative, GSH-C4, and N-acetylcysteine (NAC), which did not affect TNF-α production. The proteins identified could be useful as biomarkers of oxidative stress associated with inflammation, and further studies will be required to investigate if the extracellular forms of these proteins has immunoregulatory functions.
authors.affiliation: Institute Pasteur, Cenci-Bolognetti Foundation, "Sapienza" University of Rome, Rome, Italy; Brighton & Sussex Medical School, Falmer, Brighton, United Kingdom.
authors.forename: Paola
authors.initials: P
authors.lastname: Checconi
authors.affiliation: Brighton & Sussex Medical School, Falmer, Brighton, United Kingdom.
authors.forename: Sonia
authors.initials: S
authors.lastname: Salzano
authors.affiliation: University of Brighton, Pharmacy and Biomolecular Sciences, Moulsecoomb, Brighton, United Kingdom.
authors.forename: Lucas
authors.initials: L
authors.lastname: Bowler
authors.affiliation: Brighton & Sussex Medical School, Falmer, Brighton, United Kingdom.
authors.forename: Lisa
authors.initials: L
authors.lastname: Mullen
authors.affiliation: Brighton & Sussex Medical School, Falmer, Brighton, United Kingdom.
authors.forename: Manuela
authors.initials: M
authors.lastname: Mengozzi
authors.affiliation: Institute for Medical Biochemistry and Molecular Biology, University Medicine, Ernst-Moritz Arndt University, Greifswald, Germany.
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Hanschmann
authors.affiliation: Institute for Medical Biochemistry and Molecular Biology, University Medicine, Ernst-Moritz Arndt University, Greifswald, Germany.
authors.forename: Christopher Horst
authors.initials: CH
authors.lastname: Lillig
authors.affiliation: IRCSS San Raffaele Pisana, Telematic University, Rome, Italy.
authors.forename: Rossella
authors.initials: R
authors.lastname: Sgarbanti
authors.affiliation: IRCSS San Raffaele Pisana, Telematic University, Rome, Italy; Department of Public Health and Infectious Diseases, Institute Pasteur Cenci-Bolognetti Foundation, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Panella
authors.affiliation: Department of Public Health and Infectious Diseases, Institute Pasteur Cenci-Bolognetti Foundation, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Nencioni
authors.affiliation: IRCSS San Raffaele Pisana, Telematic University, Rome, Italy; Department of Public Health and Infectious Diseases, Institute Pasteur Cenci-Bolognetti Foundation, "Sapienza" University of Rome, Rome, Italy.
authors.forename: Anna Teresa
authors.initials: AT
authors.lastname: Palamara
authors.affiliation: Brighton & Sussex Medical School, Falmer, Brighton, United Kingdom.
authors.forename: Pietro
authors.initials: P
authors.lastname: Ghezzi
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D015153:Blotting, Western; D002460:Cell Line; D003907:Dexamethasone / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D007249:Inflammation / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D007251:Influenza, Human / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D054464:Peroxiredoxins / Q000378:metabolism; D051304:Profilins / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics; D000067996:RAW 264.7 Cells; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D013879:Thioredoxins / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism
chemical_list: D000975:Antioxidants; D008070:Lipopolysaccharides; D051304:Profilins; D011506:Proteins; D013438:Sulfhydryl Compounds; D014746:Vimentin; D013879:Thioredoxins; D003907:Dexamethasone; D054464:Peroxiredoxins; D005978:Glutathione