Cathepsin S causes inflammatory pain via biased agonism of PAR2 and TRPV4

Peishen Zhao, Tinamarie Lieu, Nicholas Barlow, Matthew Metcalf, Nicholas A. Veldhuis, Dane D. Jensen, Martina Kocan, Silvia Sostegni, Silke Haerteis, Vera Baraznenok, Ian Henderson, Erik Lindström, Raquel Guerrero-Alba, Eduardo E. Valdez-Morales, Wolfgang Liedtke, Peter McIntyre, Stephen J. Vanner, Christoph Korbmacher, Nigel W. Bunnett

Research output: Contribution to journalArticlepeer-review


Serine proteases such as trypsin and mast cell tryptase cleave protease-activated receptor-2 (PAR2) at R36↑S37and reveal a tethered ligand that excites nociceptors, causing neurogenic inflammation and pain. Whether proteases that cleave PAR2at distinct sites are biased agonists that also induce inflammation and pain is unexplored. Cathepsin S (Cat-S) is a lysosomal cysteine protease of antigen-presenting cells that is secreted during inflammation and which retains activity at extracellular pH.We observed that Cat-S cleaved PAR2at E56↑T57, which removed the canonical tethered ligand and prevented trypsin activation. In HEK and KNRK cell lines and in nociceptive neurons of mouse dorsal root ganglia, Cat-S and a decapeptide mimicking the Cat-S-revealed tethered ligand-stimulated PAR2coupling to Gαs and formation of cAMP. In contrast to trypsin, Cat-S did not mobilize intracellular Ca2+, activate ERK1/2, recruit β-arrestins, or induce PAR2endocytosis. Cat-S caused PAR2-dependent activation of transient receptor potential vanilloid 4 (TRPV4) in Xenopus laevis oocytes, HEK cells and nociceptive neurons, and stimulated neuronal hyperexcitability by adenylyl cyclase and protein kinase A-dependent mechanisms. Intraplantar injection of Cat-S caused inflammation and hyperalgesia in mice that was attenuated by PAR2or TRPV4 deletion and adenylyl cyclase inhibition. Cat-S and PAR2antagonists suppressed formalin-induced inflammation and pain, which implicates endogenous Cat-S and PAR2in inflammatory pain. Our results identify Cat-S as a biased agonist of PAR2that causes PAR2- and TRPV4-dependent inflammation and pain. They expand the role of PAR2 as a mediator of protease-driven inflammatory pain.

Original languageEnglish (US)
Pages (from-to)27215-27234
Number of pages20
JournalJournal of Biological Chemistry
Issue number39
StatePublished - Sep 26 2014

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


Dive into the research topics of 'Cathepsin S causes inflammatory pain via biased agonism of PAR2 and TRPV4'. Together they form a unique fingerprint.

Cite this