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J. Biol. Chem. 287 (41): 34101-34109

© 2012 by The American Society for Biochemistry and Molecular Biology, Inc.


Background: Cathepsin G modulates neutrophil effector functions.

Results: Cathepsin G regulates the release and proteolysis of annexin A1 and cathelin-related antimicrobial peptide, which act on neutrophil cell surface receptors to modulate the level of cellular activation.

Conclusion: Cathepsin G is required for the release of neutrophil-derived inflammatory mediators.

Significance: The unique properties of cathepsin G identify this protease as a potential therapeutic target in inflammatory processes.


Cathepsin G-regulated Release of Formyl Peptide Receptor Agonists Modulate Neutrophil Effector Functions*

Josh C. Woloszynek, Ying Hu, , and Christine T. N. Pham1

From the Department of Medicine, Division of Rheumatology, Washington University School of Medicine, St. Louis, Missouri 63110

ABSTRACT Back to Top

Abstract: Neutrophil serine proteases play an important role in inflammation by modulating neutrophil effector functions. We have previously shown that neutrophils deficient in the serine proteases cathepsin G and neutrophil elastase (CG/NE neutrophils) exhibit severe defects in chemokine CXCL2 release and reactive oxygen species (ROS) production when activated on immobilized immune complex. Exogenously added active CG rescues these defects, but the mechanism remains undefined. Using a protease-based proteomic approach, we found that, in vitro, the addition of exogenous CG to immune complex-stimulated CG/NE neutrophils led to a decrease in the level of cell-associated annexin A1 (AnxA1) and cathelin-related antimicrobial peptide (CRAMP), both known inflammatory mediators. We further confirmed that, in vivo, CG was required for the extracellular release of AnxA1 and CRAMP in a subcutaneous air pouch model. In vitro, CG efficiently cleaved AnxA1, releasing the active N-terminal peptide Ac2-26, and processed CRAMP in limited fashion. Ac2-26 and CRAMP peptides enhanced the release of CXCL2 by CG/NE neutrophils in a dose-dependent manner via formyl peptide receptor (FPR) stimulation. Blockade of FPRs by an antagonist, Boc2 (t-Boc-Phe-D-Leu-Phe-D-Leu-Phe), abrogates CXCL2 release, whereas addition of FPR agonists, fMLF and F2L, relieves Boc2 inhibition. Furthermore, the addition of active CG, but not inactive CG, also relieves Boc2 inhibition. These findings suggest that CG modulates neutrophil effector functions partly by controlling the release (and proteolysis) of FPR agonists. Unexpectedly, we found that mature CRAMP, but not Ac2-26, induced ROS production through an FPR-independent pathway.


Key Words: Chemokines • Inflammation • Neutrophil • Protease • Respiratory Burst • Formyl Peptide Receptors

Received for publication June 21, 2012. Revision received August 1, 2012.

1 To whom correspondence should be addressed: Dept. of Medicine, Div. of Rheumatology, Washington University School of Medicine, 660 South Euclid Ave., Bx. 8045, St. Louis, MO. Tel.: 314-362-9043; Fax: 314-454-1091; E-mail: cpham{at}dom.wustl.edu.



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