Eosinophil peroxidase catalyzed protein carbamylation participates in asthma

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

Research output: Contribution to journalArticle

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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 N-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.

Original languageEnglish (US)
Pages (from-to)22118-22135
Number of pages18
JournalJournal of Biological Chemistry
Volume291
Issue number42
DOIs
StatePublished - Oct 14 2016

Fingerprint

Eosinophil Peroxidase
Asthma
Proteins
Eosinophils
Post Translational Protein Processing
Phenotype
Allergens
Epithelial Cells
Scavenger Receptors
Interleukin-13
Mucins
Lysine
Chemical activation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Wang, Z., DiDonato, J. A., Buffa, J., Comhair, S. A., Aronica, M. A., Dweik, R. A., ... Hazen, S. L. (2016). Eosinophil peroxidase catalyzed protein carbamylation participates in asthma. Journal of Biological Chemistry, 291(42), 22118-22135. https://doi.org/10.1074/jbc.M116.750034

Eosinophil peroxidase catalyzed protein carbamylation participates in asthma. / Wang, Zeneng; DiDonato, Joseph A.; Buffa, Jennifer; Comhair, Suzy A.; Aronica, Mark A.; Dweik, Raed A.; Lee, Nancy A; Lee, James J.; Thomassen, Mary Jane; Kavuru, Mani; Erzurum, Serpil C.; Hazen, Stanley L.

In: Journal of Biological Chemistry, Vol. 291, No. 42, 14.10.2016, p. 22118-22135.

Research output: Contribution to journalArticle

Wang, Z, DiDonato, JA, Buffa, J, Comhair, SA, Aronica, MA, Dweik, RA, Lee, NA, Lee, JJ, Thomassen, MJ, Kavuru, M, Erzurum, SC & Hazen, SL 2016, 'Eosinophil peroxidase catalyzed protein carbamylation participates in asthma', Journal of Biological Chemistry, vol. 291, no. 42, pp. 22118-22135. https://doi.org/10.1074/jbc.M116.750034
Wang Z, DiDonato JA, Buffa J, Comhair SA, Aronica MA, Dweik RA et al. Eosinophil peroxidase catalyzed protein carbamylation participates in asthma. Journal of Biological Chemistry. 2016 Oct 14;291(42):22118-22135. https://doi.org/10.1074/jbc.M116.750034
Wang, Zeneng ; DiDonato, Joseph A. ; Buffa, Jennifer ; Comhair, Suzy A. ; Aronica, Mark A. ; Dweik, Raed A. ; Lee, Nancy A ; Lee, James J. ; Thomassen, Mary Jane ; Kavuru, Mani ; Erzurum, Serpil C. ; Hazen, Stanley L. / Eosinophil peroxidase catalyzed protein carbamylation participates in asthma. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 42. pp. 22118-22135.
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AB - 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 N∈-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.

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