Neutrophil elastase and proteinase-3 trigger G proteinbiased signaling through proteinase-activated receptor-1 (PAR1)

Authors

Koichiro Mihara, University of Calgary
Rithwik Ramachandran, University of Calgary
Bernard Renaux, University of Calgary
Mahmoud Saifeddine, University of Calgary
Morley D. Hollenberg, University of Calgary

Document Type

Article

Publication Date

11-15-2013

Journal

Journal of Biological Chemistry

Volume

288

Issue

46

First Page

32979

Last Page

32990

URL with Digital Object Identifier

10.1074/jbc.M113.483123

Abstract

Neutrophil proteinases released at sites of inflammation can affect tissue function by either activating or disarming signal transduction mediated by proteinase-activated receptors (PARs). BecausePAR1is expressed at sites where abundant neutrophil infiltration occurs, we hypothesized that neutrophil-derived enzymes might also regulate PAR1 signaling. We report here that both neutrophil elastase and proteinase-3 cleave the human PAR1 N terminus at sites distinct from the thrombin cleavage site. This cleavage results in a disarming of thrombinactivated calcium signaling through PAR1. However, the distinct non-canonical tethered ligands unmasked by neutrophil elastase and proteinase-3, as well as synthetic peptides with sequences derived from these novel exposed tethered ligands, selectively stimulated PAR1-mediated mitogen-activated protein kinase activation. This signaling was blocked by pertussis toxin, implicating a Gαi-triggered signal pathway. We conclude that neutrophil proteinases trigger biased PAR1 signaling and we describe a novel set of tethered ligands that are distinct from the classical tethered ligand revealed by thrombin. We further demonstrate the function of this biased signaling in regulating endothelial cell barrier integrity. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.