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Trypsin IV, a novel agonist of protease-activated receptors 2 and 4

Cottrell, G. S., Amadesi, S., Grady, E. F. and Bunnett, N. W. (2004) Trypsin IV, a novel agonist of protease-activated receptors 2 and 4. Journal of Biological Chemistry, 279 (14). pp. 13532-13539. ISSN 1083-351X

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To link to this item DOI: 10.1074/jbc.M312090200

Abstract/Summary

Certain serine proteases signal to cells by cleaving protease-activated receptors (PARs) and thereby regulate hemostasis, inflammation, pain and healing. However, in many tissues the proteases that activate PARs are unknown. Although pancreatic trypsin may be a physiological agonist of PAR(2) and PAR(4) in the small intestine and pancreas, these receptors are expressed by cells not normally exposed pancreatic trypsin. We investigated whether extrapancreatic forms of trypsin are PAR agonists. Epithelial cells lines from prostate, colon, and airway and human colonic mucosa expressed mRNA encoding PAR(2), trypsinogen IV, and enteropeptidase, which activates the zymogen. Immunoreactive trypsinogen IV was detected in vesicles in these cells. Trypsinogen IV was cloned from PC-3 cells and expressed in CHO cells, where it was also localized to cytoplasmic vesicles. We expressed trypsinogen IV with an N-terminal Igkappa signal peptide to direct constitutive secretion and allow enzymatic characterization. Treatment of conditioned medium with enteropeptidase reduced the apparent molecular mass of trypsinogen IV from 36 to 30 kDa and generated enzymatic activity, consistent with formation of trypsin IV. In contrast to pancreatic trypsin, trypsin IV was completely resistant to inhibition by polypeptide inhibitors. Exposure of cell lines expressing PAR(2) and PAR(4) to trypsin IV increased [Ca(2+)](i) and strongly desensitized cells to PAR agonists, whereas there were no responses in cells lacking these receptors. Thus, trypsin IV is a potential agonist of PAR(2) and PAR(4) in epithelial tissues where its resistance to endogenous trypsin inhibitors may permit prolonged signaling.

Item Type:Article
Refereed:Yes
Divisions:Faculty of Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy > Division of Pharmacology
No Reading authors. Back catalogue items
ID Code:30282
Uncontrolled Keywords:Animals Antibodies CHO Cells Caco-2 Cells Cloning, Molecular Cricetinae Enteropeptidase/genetics Epithelial Cells/cytology/enzymology Gene Expression Regulation, Enzymologic Humans Intestinal Mucosa/cytology/enzymology Isoenzymes/genetics/immunology/*metabolism Kidney/cytology Male Pancreatic Ducts/cytology Prostate/cytology Rats Receptor, PAR-2/genetics/*metabolism Receptors, Thrombin/*metabolism Trypsin/*genetics/immunology/metabolism Trypsinogen/*genetics/immunology/metabolism
Additional Information:Cottrell, Graeme S Amadesi, Silvia Grady, Eileen F Bunnett, Nigel W DK43207/DK/NIDDK NIH HHS/ DK52388/DK/NIDDK NIH HHS/ DK57840/DK/NIDDK NIH HHS/ J Biol Chem. 2004 Apr 2;279(14):13532-9. Epub 2004 Jan 15.
Publisher:American Society for Biochemistry and Molecular Biology

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