Functional study of elafin cleaved by Pseudomonas aeruginosa metalloproteinases

Nicolas Guyot, Gudmundur Bergsson, Marcus W. Butler, Catherine M. Greene, Sinéad Weldon, Efrat Kessler, Rodney L. Levine, Shane J. O'Neill, Clifford C. Taggart, Noel G. McElvaney

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Elafin is a 6-kDa innate immune protein present at several epithelial surfaces including the pulmonary epithelium. It is a canonical protease inhibitor of two neutrophil serine proteases wneutrophil elastase (NE) and proteinase 3x with the capacity to covalently bind extracellular matrix proteins by transglutamination. In addition to these properties, elafin also possesses antimicrobial and immunomodulatory activities. The aim of the present study was to investigate the effect of Pseudomonas aeruginosa proteases on elafin function. We found that P. aeruginosa PAO1-conditioned medium and two purified Pseudomonas metalloproteases, pseudolysin (elastase) and aeruginolysin (alkaline protease), are able to cleave recombinant elafin. Pseudolysin was shown to inactivate the anti-NE activity of elafin by cleaving its protease-binding loop. Interestingly, antibacterial properties of elafin against PAO1 were found to be unaffected after pseudolysin treatment. In contrast to pseudolysin, aeruginolysin failed to inactivate the inhibitory properties of elafin against NE. Aeruginolysin cleaves elafin at the amino-terminal Lys6- Gly7 peptide bond, resulting in a decreased ability to covalently bind purified fibronectin following transglutaminase activity. In conclusion, this study provides evidence that elafin is susceptible to proteolytic cleavage at alternative sites by P. aeruginosa metalloproteinases, which can affect different biological functions of elafin.

Original languageEnglish
Pages (from-to)705-716
Number of pages12
JournalBiological Chemistry
Volume391
Issue number6
DOIs
StatePublished - Jun 2010

Funding

FundersFunder number
Alpha One Foundation
Cystic Fibrosis Association of Ireland
Cystic Fibrosis Hope Source
Cystic Fibrosis Research Trust
National Heart, Lung, and Blood InstituteZIAHL000225
Health Research Board
Royal College of Surgeons in Ireland
Higher Education Authority
Science Foundation Ireland

    Keywords

    • Antimicrobials
    • Antiproteases
    • Cleavage
    • Microbial
    • Proteases
    • Transglutaminase

    Fingerprint

    Dive into the research topics of 'Functional study of elafin cleaved by Pseudomonas aeruginosa metalloproteinases'. Together they form a unique fingerprint.

    Cite this