Chitinase activates protease-activated receptor-2 in human airway epithelial cells

Hee Hong Jeong, Yeon Hong Jung, Boryung Park, Syng Ill Lee, Taeg Seo Jeong, Kyu Earn Kim, Hyun Sohn Myung, DongMin Shin

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Mammalian chitinase released by airway epithelia is thought to be an important mediator of disease manifestation in an experimental model of asthma. However, the intracellular signaling mechanisms engaged by exogenous chitinase in human airway epithelial cells are unknown. Here, we investigated the direct effects of exogenous chitinase from Streptomyces griseus on Ca 2+ signaling in human airway epithelial cells. Spectrofluorometry was used to measure intracellular Ca 2+ concentration ([Ca 2+ ] i ) in fura-2-AM-loaded cells. S. griseus chitinase induced dose-dependent [Ca 2+ ] i increases in normal human bronchial epithelial cells and promoted [Ca 2+ ] i oscillations in H292 cells. Chitinase-induced [Ca 2+ ] i oscillations were independent of extracellular Ca 2+ , suggesting that the observed [Ca 2+ ] i increases were due to Ca 2+ release from intracellular stores. Accordingly, after depleting endoplasmic reticulum (ER) Ca 2+ with the ER Ca 2+ ATPase inhibitor, thapsigargin, chitinase-mediated [Ca 2+ ] i increases were abolished. Treatment with the phospholipase C (PLC) inhibitor U73122 or the 1, 4, 5-trisinositolphosphate (IP 3 ) receptor inhibitor 2-APB attenuated chitinase-induced [Ca 2+ ] i increases. Desensitization of protease-activated receptor-2 (PAR-2) by repetitive agonist stimulation or siRNA-mediated PAR-2 knockdown revealed that chitinase-mediated [Ca 2+ ] i increases were exclusively mediated by PAR-2 activation. Finally, chitinase was found to cleave a model peptide representing the cleavage site of PAR-2 and enhanced IL-8 production. These results indicate that exogenous chitinase is a potent proteolytic activator of PAR-2 that can directly induce PLC/IP 3 -dependent Ca 2+ signaling in human airway epithelial cells.

Original languageEnglish
Pages (from-to)530-535
Number of pages6
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume39
Issue number5
DOIs
Publication statusPublished - 2008 Nov 1

Fingerprint

PAR-2 Receptor
Chitinases
Epithelial Cells
Streptomyces griseus
Type C Phospholipases
Endoplasmic Reticulum
Thapsigargin
Fura-2
Fluorescence Spectrometry
Interleukin-8
Small Interfering RNA
Adenosine Triphosphatases
Theoretical Models
Asthma
Epithelium
Chemical activation

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

Jeong, Hee Hong ; Jung, Yeon Hong ; Park, Boryung ; Lee, Syng Ill ; Jeong, Taeg Seo ; Kim, Kyu Earn ; Myung, Hyun Sohn ; Shin, DongMin. / Chitinase activates protease-activated receptor-2 in human airway epithelial cells. In: American Journal of Respiratory Cell and Molecular Biology. 2008 ; Vol. 39, No. 5. pp. 530-535.
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Chitinase activates protease-activated receptor-2 in human airway epithelial cells. / Jeong, Hee Hong; Jung, Yeon Hong; Park, Boryung; Lee, Syng Ill; Jeong, Taeg Seo; Kim, Kyu Earn; Myung, Hyun Sohn; Shin, DongMin.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 39, No. 5, 01.11.2008, p. 530-535.

Research output: Contribution to journalArticle

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