Eosinophil Peroxidase Catalyzed Protein Carbamylation Participates in Asthma.

Abstract

The biochemical mechanisms through which eosinophils contribute to asthma pathogenesis are unclear. Here we show eosinophil peroxidase (EPO), an abundant granule protein released by activated eosinophils, contributes to characteristic asthma-related phenotypes through oxidative posttranslational modification (PTM) of proteins in asthmatic airways through a process called carbamylation. Using a combination of studies we now show EPO uses plasma levels of the pseudohalide thiocyanate (SCN-) as substrate to catalyze protein carbamylation, as monitored by PTM of protein lysine residues into Nϵ-carbamyllysine (homocitrulline), and contributes to the pathophysiological sequelae of eosinophil activation. Studies using EPO-deficient mice confirm EPO serves as a major enzymatic source for protein carbamylation during eosinophilic inflammatory models, including aeroallergen challenge. Clinical studies similarly revealed significant enrichment in carbamylation of airway proteins recovered from atopic asthmatics versus healthy controls in response to segmental allergen challenge. Protein-bound homocitrulline is shown to be co-localized with EPO within human asthmatic airways. Moreover, pathophysiologically relevant levels of carbamylated protein either incubated with cultured human airway epithelial cells in vitro, or provided as an aerosolized exposure in non-sensitized mice, induced multiple asthma-associated phenotypes including induction of mucin, Th2 cytokines, IFNγ, TGFβ, and epithelial cell apoptosis. Studies with scavenger receptor-A1 null mice reveal reduced IL-13 generation following exposure to aerosolized carbamylated protein, but no changes in other asthma-related phenotypes. In summary, EPO-mediated protein carbamylation is promoted during allergen-induced asthma exacerbation, and can both modulate immune responses and trigger a cascade of many of the inflammatory signals present in asthma.

• Authors: Zeneng Wang, Joseph A DiDonato, Jennifer Buffa, Suzy A Comhair, Mark A Aronica, Raed A Dweik, Nancy A Lee, James J Lee, Mary Jane Thomassen, Mani Kavuru, Serpil C Erzurum, Stanley L Hazen
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 291 Issue 42 Pg. 22118-22135 (Oct 14 2016) ISSN: 1083-351X [Electronic] United States
• PMID: 27587397 (Publication Type: Journal Article)
• Copyright: © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Chemical References

• IFNG protein, human
• IFNG protein, mouse
• Interleukin-13
• Transforming Growth Factor beta
• homocitrulline
• Citrulline
• Interferon-gamma
• Eosinophil Peroxidase

Topics

• A549 Cells
• Animals
• Asthma (immunology, pathology)
• Citrulline (analogs & derivatives, immunology)
• Eosinophil Peroxidase (immunology)
• Eosinophils (immunology, pathology)
• Humans
• Interferon-gamma (immunology)
• Interleukin-13 (immunology)
• Mice
• Protein Processing, Post-Translational (immunology)
• Th2 Cells (immunology, pathology)
• Transforming Growth Factor beta (immunology)