JPET Introducing ALZET?ew Model 2006 Pump

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

Journal of Pharmacology And Experimental Therapeutics Fast Forward
First published on November 25, 2002; DOI: 10.1124/jpet.102.043844

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
jpet.102.043844v1
304/3/1120    most recent
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Al-Ani, B.
Right arrow Articles by Hollenberg, M. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Al-Ani, B.
Right arrow Articles by Hollenberg, M. D.
Right arrowPubmed/NCBI databases
*Gene*GEO Profiles
*HomoloGene*Nucleotide
*Protein*UniGene
*Compound via MeSH
*Substance via MeSH

Vol. 304, Issue 3, 1120-1128, March 2003

Selective Tryptic Cleavage at the Tethered Ligand Site of the Amino Terminal Domain of Proteinase-Activated Receptor-2 in Intact Cells

Bahjat Al-Ani and Morley D. Hollenberg

Diabetes/Endocrine and Mucosal Inflammation Research Groups, Departments of Pharmacology and Therapeutics (B.A.-A., M.D.H.) and Medicine (M.D.H.), University of Calgary Faculty of Medicine, Calgary, Alberta, Canada

In intact cells, trypsin activates proteinase-activated receptor-2 (PAR2) by hydrolysis at residues R36/S37 (amino acids are abbreviated by their one-letter code), revealing an active tethered ligand sequence. We sought to determine whether in intact cells, the tryptic cleavage/activation of PAR2 might also be accompanied by hydrolysis at other potential N-terminal cleavage sites, like residues K34, R41, K51, and K72, as implied by the tryptic cleavage in vitro at these residues of Escherichia coli-expressed human N-terminal PAR2R31-P79. To this end, four PAR2 mutants with altered tryptic cleavage sites were prepared (PAR2R36A, PAR2S37P, PAR2R41A, and PAR2R36AR41A), expressed in Kirsten virus-transformed rat kidney cells and were evaluated together with the wild-type PAR2-expressing cells for 1) activation (Ca2+ signaling) by trypsin and the receptor-activating peptide SLIGRL-NH2 (SL-NH2) and 2) the tryptic release of two antigenic receptor determinants, one N-terminal to the R36/S37 cleavage/activation site detected by SLAW-A antibody and the second (detected by antibody, B5), N-terminal to residues K51, K72. None of the mutants resistant to cleavage at R36 were activated by trypsin, yet all retained reactivity to B5 and all were activated by SL-NH2. In contrast, trypsin activated both wild-type and PAR2R41A, leading to a disappearance of SLAW-A but not B5 reactivity. We conclude that, as opposed to the E. coli-expressed PAR2 N-terminal polypeptide, PAR2 expressed in intact cells displays selective tryptic cleavage at the R36/S37 activation site, without cleaving downstream. Thus, in intact cells, trypsin activation does not concurrently "disarm" rat PAR2, but leaves the "tethered ligand" persistently attached to the body of the receptor.

0022-3565/03/3043-1120$07.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2003 by The American Society for Pharmacology and Experimental Therapeutics


This article has been cited by other articles:


Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
M. Saifeddine, M. L. Seymour, Y.-P. Xiao, S. J. Compton, S. Houle, R. Ramachandran, W. K. MacNaughton, S. Simonet, C. Vayssettes-Courchay, T. J. Verbeuren, et al.
Proteinase-activated receptor-2 activating peptides: distinct canine coronary artery receptor systems
Am J Physiol Heart Circ Physiol, December 1, 2007; 293(6): H3279 - H3289.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
T. Nishiyama, T. Nakamura, K. Obara, H. Inoue, K. Mishima, N. Matsumoto, M. Matsui, T. Manabe, K. Mikoshiba, and I. Saito
Up-Regulated PAR-2-Mediated Salivary Secretion in Mice Deficient in Muscarinic Acetylcholine Receptor Subtypes
J. Pharmacol. Exp. Ther., February 1, 2007; 320(2): 516 - 524.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
K. Oikonomopoulou, K. K. Hansen, M. Saifeddine, I. Tea, M. Blaber, S. I. Blaber, I. Scarisbrick, P. Andrade-Gordon, G. S. Cottrell, N. W. Bunnett, et al.
Proteinase-activated Receptors, Targets for Kallikrein Signaling
J. Biol. Chem., October 27, 2006; 281(43): 32095 - 32112.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
E. Adam, K. K. Hansen, O. F. Astudillo, L. Coulon, F. Bex, X. Duhant, E. Jaumotte, M. D. Hollenberg, and A. Jacquet
The House Dust Mite Allergen Der p 1, Unlike Der p 3, Stimulates the Expression of Interleukin-8 in Human Airway Epithelial Cells via a Proteinase-activated Receptor-2-independent Mechanism
J. Biol. Chem., March 17, 2006; 281(11): 6910 - 6923.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Respir. Cell Mol. Bio.Home page
S. Dulon, D. Leduc, G. S. Cottrell, J. D'Alayer, K. K. Hansen, N. W. Bunnett, M. D. Hollenberg, D. Pidard, and M. Chignard
Pseudomonas aeruginosa Elastase Disables Proteinase-Activated Receptor 2 in Respiratory Epithelial Cells
Am. J. Respir. Cell Mol. Biol., May 1, 2005; 32(5): 411 - 419.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
B. Al-Ani, K. K. Hansen, and M. D. Hollenberg
Proteinase-Activated Receptor-2: Key Role of Amino-Terminal Dipeptide Residues of the Tethered Ligand for Receptor Activation
Mol. Pharmacol., January 1, 2004; 65(1): 149 - 156.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
All ASPET Journals Molecular Pharmacology Pharmacological Reviews
Molecular Interventions Drug Metabolism and Disposition

Copyright © 2003 by the American Society for Pharmacology and Experimental Therapeutics.