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Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors 1 and 2 to induce inflammation and hyperalgesia

Knecht, Wolfgang LU ; Cottrell, Graeme S ; Amadesi, Silvia ; Mohlin, Johanna ; Skåregärde, Anita ; Gedda, Karin ; Peterson, Anders ; Chapman, Kevin ; Hollenberg, Morley D and Vergnolle, Nathalie , et al. (2007) In The Journal of biological chemistry 282(36). p.26089-26100
Abstract

Although principally produced by the pancreas to degrade dietary proteins in the intestine, trypsins are also expressed in the nervous system and in epithelial tissues, where they have diverse actions that could be mediated by protease-activated receptors (PARs). We examined the biological actions of human trypsin IV (or mesotrypsin) and rat p23, inhibitor-resistant forms of trypsin. The zymogens trypsinogen IV and pro-p23 were expressed in Escherichia coli and purified to apparent homogeneity. Enteropeptidase cleaved both zymogens, liberating active trypsin IV and p23, which were resistant to soybean trypsin inhibitor and aprotinin. Trypsin IV cleaved N-terminal fragments of PAR(1), PAR(2), and PAR(4) at sites that would expose the... (More)

Although principally produced by the pancreas to degrade dietary proteins in the intestine, trypsins are also expressed in the nervous system and in epithelial tissues, where they have diverse actions that could be mediated by protease-activated receptors (PARs). We examined the biological actions of human trypsin IV (or mesotrypsin) and rat p23, inhibitor-resistant forms of trypsin. The zymogens trypsinogen IV and pro-p23 were expressed in Escherichia coli and purified to apparent homogeneity. Enteropeptidase cleaved both zymogens, liberating active trypsin IV and p23, which were resistant to soybean trypsin inhibitor and aprotinin. Trypsin IV cleaved N-terminal fragments of PAR(1), PAR(2), and PAR(4) at sites that would expose the tethered ligand (PAR(1) = PAR(4) > PAR(2)). Trypsin IV increased [Ca(2+)](i) in transfected cells expressing human PAR(1) and PAR(2) with similar potencies (PAR(1), 0.5 microm; PAR(2), 0.6 microm). p23 also cleaved fragments of PAR(1) and PAR(2) and signaled to cells expressing these receptors. Trypsin IV and p23 increased [Ca(2+)](i) in rat dorsal root ganglion neurons that responded to capsaicin and which thus mediate neurogenic inflammation and nociception. Intraplantar injection of trypsin IV and p23 in mice induced edema and granulocyte infiltration, which were not observed in PAR (-/-)(1)(trypsin IV) and PAR (-/-)(2) (trypsin IV and p23) mice. Trypsin IV and p23 caused thermal hyperalgesia and mechanical allodynia and hyperalgesia in mice, and these effects were absent in PAR (-/-)(2) mice but maintained in PAR (-/-)(1) mice. Thus, trypsin IV and p23 are inhibitor-resistant trypsins that can cleave and activate PARs, causing PAR(1)- and PAR(2)-dependent inflammation and PAR(2)-dependent hyperalgesia.

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type
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publication status
published
keywords
Animals, Aprotinin/chemistry, Calcium Signaling/drug effects, Capsaicin/pharmacology, Edema/chemically induced, Enteropeptidase/chemistry, Ganglia, Spinal/metabolism, Granulocytes/metabolism, Humans, Hyperalgesia/chemically induced, Inflammation/chemically induced, Male, Mice, Mice, Knockout, Nociceptors/metabolism, Pain Measurement, Rats, Rats, Sprague-Dawley, Receptor, PAR-1/deficiency, Receptor, PAR-2/deficiency, Receptors, Proteinase-Activated/metabolism, Receptors, Thrombin/metabolism, Recombinant Proteins/chemistry, Trypsin/chemistry, Trypsin Inhibitors/chemistry
in
The Journal of biological chemistry
volume
282
issue
36
pages
26089 - 26100
publisher
American Society for Biochemistry and Molecular Biology
external identifiers
  • scopus:34548829096
  • pmid:17623652
ISSN
0021-9258
DOI
10.1074/jbc.M703840200
language
English
LU publication?
no
id
1e87d02f-6bf5-461b-bf6e-6035419e335a
date added to LUP
2020-07-22 14:10:24
date last changed
2024-01-02 14:59:34
@article{1e87d02f-6bf5-461b-bf6e-6035419e335a,
  abstract     = {{<p>Although principally produced by the pancreas to degrade dietary proteins in the intestine, trypsins are also expressed in the nervous system and in epithelial tissues, where they have diverse actions that could be mediated by protease-activated receptors (PARs). We examined the biological actions of human trypsin IV (or mesotrypsin) and rat p23, inhibitor-resistant forms of trypsin. The zymogens trypsinogen IV and pro-p23 were expressed in Escherichia coli and purified to apparent homogeneity. Enteropeptidase cleaved both zymogens, liberating active trypsin IV and p23, which were resistant to soybean trypsin inhibitor and aprotinin. Trypsin IV cleaved N-terminal fragments of PAR(1), PAR(2), and PAR(4) at sites that would expose the tethered ligand (PAR(1) = PAR(4) &gt; PAR(2)). Trypsin IV increased [Ca(2+)](i) in transfected cells expressing human PAR(1) and PAR(2) with similar potencies (PAR(1), 0.5 microm; PAR(2), 0.6 microm). p23 also cleaved fragments of PAR(1) and PAR(2) and signaled to cells expressing these receptors. Trypsin IV and p23 increased [Ca(2+)](i) in rat dorsal root ganglion neurons that responded to capsaicin and which thus mediate neurogenic inflammation and nociception. Intraplantar injection of trypsin IV and p23 in mice induced edema and granulocyte infiltration, which were not observed in PAR (-/-)(1)(trypsin IV) and PAR (-/-)(2) (trypsin IV and p23) mice. Trypsin IV and p23 caused thermal hyperalgesia and mechanical allodynia and hyperalgesia in mice, and these effects were absent in PAR (-/-)(2) mice but maintained in PAR (-/-)(1) mice. Thus, trypsin IV and p23 are inhibitor-resistant trypsins that can cleave and activate PARs, causing PAR(1)- and PAR(2)-dependent inflammation and PAR(2)-dependent hyperalgesia.</p>}},
  author       = {{Knecht, Wolfgang and Cottrell, Graeme S and Amadesi, Silvia and Mohlin, Johanna and Skåregärde, Anita and Gedda, Karin and Peterson, Anders and Chapman, Kevin and Hollenberg, Morley D and Vergnolle, Nathalie and Bunnett, Nigel W}},
  issn         = {{0021-9258}},
  keywords     = {{Animals; Aprotinin/chemistry; Calcium Signaling/drug effects; Capsaicin/pharmacology; Edema/chemically induced; Enteropeptidase/chemistry; Ganglia, Spinal/metabolism; Granulocytes/metabolism; Humans; Hyperalgesia/chemically induced; Inflammation/chemically induced; Male; Mice; Mice, Knockout; Nociceptors/metabolism; Pain Measurement; Rats; Rats, Sprague-Dawley; Receptor, PAR-1/deficiency; Receptor, PAR-2/deficiency; Receptors, Proteinase-Activated/metabolism; Receptors, Thrombin/metabolism; Recombinant Proteins/chemistry; Trypsin/chemistry; Trypsin Inhibitors/chemistry}},
  language     = {{eng}},
  month        = {{09}},
  number       = {{36}},
  pages        = {{26089--26100}},
  publisher    = {{American Society for Biochemistry and Molecular Biology}},
  series       = {{The Journal of biological chemistry}},
  title        = {{Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors 1 and 2 to induce inflammation and hyperalgesia}},
  url          = {{http://dx.doi.org/10.1074/jbc.M703840200}},
  doi          = {{10.1074/jbc.M703840200}},
  volume       = {{282}},
  year         = {{2007}},
}