title: Crotalphine desensitizes TRPA1 ion channels to alleviate inflammatory hyperalgesia.
abstract: Crotalphine is a structural analogue to a novel analgesic peptide that was first identified in the crude venom from the South American rattlesnake Crotalus durissus terrificus. Although crotalphine's analgesic effect is well established, its direct mechanism of action remains unresolved. The aim of the present study was to investigate the effect of crotalphine on ion channels in peripheral pain pathways. We found that picomolar concentrations of crotalphine selectively activate heterologously expressed and native TRPA1 ion channels. TRPA1 activation by crotalphine required intact N-terminal cysteine residues and was followed by strong and long-lasting desensitization of the channel. Homologous desensitization of recombinant TRPA1 and heterologous desensitization in cultured dorsal root ganglia neurons was observed. Likewise, crotalphine acted on peptidergic TRPA1-expressing nerve endings ex vivo as demonstrated by suppression of calcitonin gene-related peptide release from the trachea and in vivo by inhibition of chemically induced and inflammatory hypersensitivity in mice. The crotalphine-mediated desensitizing effect was abolished by the TRPA1 blocker HC030031 and absent in TRPA1-deficient mice. Taken together, these results suggest that crotalphine is the first peptide to mediate antinociception selectively and at subnanomolar concentrations by targeting TRPA1 ion channels.
authors.affiliation: Institute of Physiology and Pathophysiology, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.|Department of Physiology, University Hospital RWTH Aachen, Aachen, Germany.
authors.forename: Elisangela
authors.initials: E
authors.lastname: Bressan
authors.affiliation: Institute of Physiology and Pathophysiology, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.|Department of Cellular Neurophysiology, Institute of Physiology of the Czech Academy of Sciences, Prague, Czech Republic.
authors.forename: Filip
authors.initials: F
authors.lastname: Touska
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, St. Lucia, Australia.
authors.forename: Irina
authors.initials: I
authors.lastname: Vetter
authors.affiliation: Institute of Physiology and Pathophysiology, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Kistner
authors.affiliation: Institute of Physiology and Pathophysiology, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.
authors.forename: Tatjana I
authors.initials: TI
authors.lastname: Kichko
authors.affiliation: Laboratory of Pain and Signaling, Butantan Institute, São Paulo, Brazil.
authors.forename: Nathália B
authors.initials: NB
authors.lastname: Teixeira
authors.affiliation: Laboratory of Pain and Signaling, Butantan Institute, São Paulo, Brazil.
authors.forename: Gisele
authors.initials: G
authors.lastname: Picolo
authors.affiliation: Laboratory of Pain and Signaling, Butantan Institute, São Paulo, Brazil.
authors.forename: Yara
authors.initials: Y
authors.lastname: Cury
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, St. Lucia, Australia.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Lewis
authors.affiliation: Institute of Physiology and Pathophysiology, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.
authors.forename: Michael J M
authors.initials: MJM
authors.lastname: Fischer
authors.affiliation: Institute of Physiology and Pathophysiology, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.|Department of Anesthesiology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Zimmermann
authors.affiliation: Institute of Physiology and Pathophysiology, Friedrich-Alexander University Erlangen-Nurnberg, Erlangen, Germany.
authors.forename: Peter W
authors.initials: PW
authors.lastname: Reeh
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000700:Analgesics / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D001920:Bradykinin / Q000633:toxicity; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005727:Ganglia, Spinal / Q000166:cytology; D057809:HEK293 Cells; D006801:Humans; D006930:Hyperalgesia / Q000139:chemically induced / Q000188:drug therapy* / Q000209:etiology; D007249:Inflammation / Q000150:complications; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology / Q000627:therapeutic use*; D000074025:TRPA1 Cation Channel; D050051:Transient Receptor Potential Channels / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015054:Zymosan / Q000633:toxicity
chemical_list: D000700:Analgesics; D010455:Peptides; D000074025:TRPA1 Cation Channel; D050051:Transient Receptor Potential Channels; C475685:Trpa1 protein, mouse; C533006:crotalphine; D015054:Zymosan; D001920:Bradykinin; D002118:Calcium