Collagenolytic matrix metalloproteinases antagonize proteinase-activated receptor-2 activation, providing insights into extracellular matrix turnover



Falconer, Adrian MD, Chan, Chun Ming, Gray, Joseph, Nagashima, Izuru, Holland, Robert A, Shimizu, Hiroki, Pickford, Andrew R, Rowan, Andrew D and Wilkinson, David J ORCID: 0000-0002-6262-3575
(2019) Collagenolytic matrix metalloproteinases antagonize proteinase-activated receptor-2 activation, providing insights into extracellular matrix turnover. JOURNAL OF BIOLOGICAL CHEMISTRY, 294 (26). pp. 10266-10277.

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Abstract

The collagenase subfamily of matrix metalloproteinases (MMPs) have important roles in the remodeling of collagenous matrices. The proteinase-activated receptor (PAR) family has a unique mechanism of activation requiring proteolysis of an extracellular domain forming a neo-N terminus that acts as a tethered ligand, a process that has been associated with the development of arthritis. Canonical PAR2 activation typically occurs via a serine proteinase at Arg<sup>36</sup>-Ser<sup>37</sup>, but other proteinases can cleave PARs downstream of the tethered ligand and "disarm" the receptor. To identify additional cleavage sites within PAR2, we synthesized a 42-amino-acid peptide corresponding to the extracellular region. We observed that all three soluble MMP collagenases, MMP-1, MMP-8, and MMP-13, cleave PAR2 and discovered a novel cleavage site (Ser<sup>37</sup>-Leu<sup>38</sup>). Metalloproteinases from resorbing bovine nasal cartilage and recombinant human collagenases could cleave a quenched fluorescent peptide mimicking the canonical PAR2 activation region, and kinetic constants were determined. In PAR2-overexpressing SW1353 chondrocytes, we demonstrated that the activator peptide SLIGKV-NH<sub>2</sub> induces rapid calcium flux, inflammatory gene expression (including <i>MMP1</i> and <i>MMP13</i>), and the phosphorylation of extracellular signal-regulated kinase (ERK) and p38 kinase. The corresponding MMP cleavage-derived peptide (LIGKVD-NH<sub>2</sub>) exhibited no canonical activation; however, we observed phosphorylation of ERK, providing evidence of biased agonism. Importantly, we demonstrated that preincubation with active MMP-1 reduced downstream PAR2 activation by a canonical activator, matriptase, but not SLIGKV-NH<sub>2</sub> These results support a role for collagenases as proteinases capable of disarming PAR2, revealing a mechanism that suppresses PAR2-mediated inflammatory responses.

Item Type: Article
Uncontrolled Keywords: matrix metalloproteinase (MMP), cell signaling, proteolysis, chondrocyte, arthritis, extracellular matrix, inflammation, collagenase, proteinase-activated receptor-2 (PAR2)
Depositing User: Symplectic Admin
Date Deposited: 27 Aug 2019 14:28
Last Modified: 19 Jan 2023 00:28
DOI: 10.1074/jbc.RA119.006974
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3052646