Trypsin IV or mesotrypsin and p23 cleave protease-activated receptors 1 and 2 to induce inflammation and hyperalgesia.
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.
Knecht
Wolfgang
W
Molecular Pharmacology and Lead Generation, AstraZeneca Research and Development, Mölndal 431 83, Sweden.
Cottrell
Graeme S
GS
Amadesi
Silvia
S
Mohlin
Johanna
J
Skåregärde
Anita
A
Gedda
Karin
K
Peterson
Anders
A
Chapman
Kevin
K
Hollenberg
Morley D
MD
Vergnolle
Nathalie
N
Bunnett
Nigel W
NW
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
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