title: Mechanisms of transient receptor potential vanilloid 1 activation and sensitization by allyl isothiocyanate.
abstract: Allyl isothiocyanate (AITC; aka, mustard oil) is a powerful irritant produced by Brassica plants as a defensive trait against herbivores and confers pungency to mustard and wasabi. AITC is widely used experimentally as an inducer of acute pain and neurogenic inflammation, which are largely mediated by the activation of nociceptive cation channels transient receptor potential ankyrin 1 and transient receptor potential vanilloid 1 (TRPV1). Although it is generally accepted that electrophilic agents activate these channels through covalent modification of cytosolic cysteine residues, the mechanism underlying TRPV1 activation by AITC remains unknown. Here we show that, surprisingly, AITC-induced activation of TRPV1 does not require interaction with cysteine residues, but is largely dependent on S513, a residue that is involved in capsaicin binding. Furthermore, AITC acts in a membrane-delimited manner and induces a shift of the voltage dependence of activation toward negative voltages, which is reminiscent of capsaicin effects. These data indicate that AITC acts through reversible interactions with the capsaicin binding site. In addition, we show that TRPV1 is a locus for cross-sensitization between AITC and acidosis in nociceptive neurons. Furthermore, we show that residue F660, which is known to determine the stimulation by low pH in human TRPV1, is also essential for the cross-sensitization of the effects of AITC and low pH. Taken together, these findings demonstrate that not all reactive electrophiles stimulate TRPV1 via cysteine modification and help understanding the molecular bases underlying the surprisingly large role of this channel as mediator of the algesic properties of AITC.
authors.affiliation: Laboratory for Ion Channel Research, Department of Molecular Cell Biology and TRP Research Platform Leuven-TRPLe, KU Leuven, Leuven, Belgium.
authors.forename: Maarten
authors.initials: M
authors.lastname: Gees
authors.forename: Yeranddy A
authors.initials: YA
authors.lastname: Alpizar
authors.forename: Brett
authors.initials: B
authors.lastname: Boonen
authors.forename: Alicia
authors.initials: A
authors.lastname: Sanchez
authors.forename: Wouter
authors.initials: W
authors.lastname: Everaerts
authors.forename: Andrei
authors.initials: A
authors.lastname: Segal
authors.forename: Fenqin
authors.initials: F
authors.lastname: Xue
authors.forename: Annelies
authors.initials: A
authors.lastname: Janssens
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Owsianik
authors.forename: Bernd
authors.initials: B
authors.lastname: Nilius
authors.forename: Thomas
authors.initials: T
authors.lastname: Voets
authors.forename: Karel
authors.initials: K
authors.lastname: Talavera
mesh_terms: D000818:Animals; D001665:Binding Sites; D002211:Capsaicin / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D017879:Isothiocyanates / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D018408:Patch-Clamp Techniques; D011984:Sensory Receptor Cells / Q000187:drug effects / Q000378:metabolism; D050916:TRPV Cation Channels / Q000819:agonists / Q000235:genetics / Q000378:metabolism*
chemical_list: D017879:Isothiocyanates; D050916:TRPV Cation Channels; C494484:TRPV1 receptor; C004471:allyl isothiocyanate; D002211:Capsaicin