title: Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors 1 and 2 to induce inflammation and hyperalgesia.
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 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.
authors.affiliation: Molecular Pharmacology and Lead Generation, AstraZeneca Research and Development, Mölndal 431 83, Sweden.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Knecht
authors.forename: Graeme S
authors.initials: GS
authors.lastname: Cottrell
authors.forename: Silvia
authors.initials: S
authors.lastname: Amadesi
authors.forename: Johanna
authors.initials: J
authors.lastname: Mohlin
authors.forename: Anita
authors.initials: A
authors.lastname: Skåregärde
authors.forename: Karin
authors.initials: K
authors.lastname: Gedda
authors.forename: Anders
authors.initials: A
authors.lastname: Peterson
authors.forename: Kevin
authors.initials: K
authors.lastname: Chapman
authors.forename: Morley D
authors.initials: MD
authors.lastname: Hollenberg
authors.forename: Nathalie
authors.initials: N
authors.lastname: Vergnolle
authors.forename: Nigel W
authors.initials: NW
authors.lastname: Bunnett
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000737:chemistry; D020013:Calcium Signaling / Q000187:drug effects; D002211:Capsaicin / Q000494:pharmacology; D004487:Edema / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004765:Enteropeptidase / Q000737:chemistry; D005727:Ganglia, Spinal / Q000378:metabolism / Q000473:pathology; D006098:Granulocytes / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006930:Hyperalgesia / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009619:Nociceptors / Q000378:metabolism / Q000473:pathology; D010147:Pain Measurement; D051381:Rats; D017207:Rats, Sprague-Dawley; D044463:Receptor, PAR-1 / Q000172:deficiency / Q000378:metabolism*; D044464:Receptor, PAR-2 / Q000172:deficiency / Q000502:physiology*; D044462:Receptors, Proteinase-Activated / Q000378:metabolism; D018179:Receptors, Thrombin / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D014357:Trypsin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D014361:Trypsin Inhibitors / Q000737:chemistry
chemical_list: D044463:Receptor, PAR-1; D044464:Receptor, PAR-2; D044462:Receptors, Proteinase-Activated; D018179:Receptors, Thrombin; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors; C481942:protease-activated receptor 4, mouse; D007611:Aprotinin; C040749:PRSS3 protein, human; D014357:Trypsin; D004765:Enteropeptidase; C112908:protease-activated receptor 4; D002211:Capsaicin