title: The pore properties of human nociceptor channel TRPA1 evaluated in single channel recordings.
abstract: TRPA channels detect stimuli of different sensory modalities, including a broad spectrum of chemosensory stimuli, noxious stimuli associated with tissue damage and inflammation, mechanical stimuli, and thermal stimuli. Despite a growing understanding of potential modulators, agonists, and antagonists for these channels, the exact mechanisms of channel regulation and activation remain mostly unknown or controversial and widely debated. Relatively little is also known about the basic biophysical parameters of both native and heterologously expressed TRPA channels. Here we use conventional single channel inside-out and outside-out patch recording from the human TRPA1 channel transiently expressed in human embryonic kidney 293T cells to characterize the selectivity of the channel for inorganic mono-/divalent and organic monovalent cations in the presence of allylisothiocyanate (AITC). We show the relative permeability of the hTRPA1 channel to inorganic cations to be:and to organic cations:Na(+)(1.0)≥ dimethylamine (0.99)>trimethylamine (0.7)>tetramethylammonium (0.4)>N-methyl-d-glucamine (0.1). Activation of the hTRPA1 channels by AITC appears to recruit the channels to a conformational state with an increased permeability to large organic cations. The pore of the channels in this state can be characterized as dilated by approximately 1-2.5 Å. These findings provide important insight into the basic fundamental properties and function of TRPA1 channels in general and human TRPA1 channel in particular.
authors.affiliation: Whitney Laboratory, Center for Smell and Taste, St. Augustine, FL, USA. bobkov@whitney.ufl.edu
authors.forename: Y V
authors.initials: YV
authors.lastname: Bobkov
authors.forename: E A
authors.initials: EA
authors.lastname: Corey
authors.forename: B W
authors.initials: BW
authors.lastname: Ache
mesh_terms: D002118:Calcium / Q000494:pharmacology; D015220:Calcium Channels / Q000235:genetics / Q000502:physiology*; D002413:Cations, Divalent / Q000494:pharmacology; D002414:Cations, Monovalent / Q000494:pharmacology; D004123:Dimethylamines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D057809:HEK293 Cells; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology*; D017879:Isothiocyanates / Q000494:pharmacology; D008536:Meglumine / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects; D008744:Methylamines / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000502:physiology*; D009619:Nociceptors / Q000502:physiology; D018408:Patch-Clamp Techniques; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D012430:Ruthenium Red / Q000494:pharmacology; D000074025:TRPA1 Cation Channel; D014162:Transfection; D050051:Transient Receptor Potential Channels / Q000235:genetics / Q000502:physiology*
chemical_list: D015220:Calcium Channels; D002413:Cations, Divalent; D002414:Cations, Monovalent; D004123:Dimethylamines; D017879:Isothiocyanates; D008744:Methylamines; D009419:Nerve Tissue Proteins; D000644:Quaternary Ammonium Compounds; D000074025:TRPA1 Cation Channel; C473350:TRPA1 protein, human; D050051:Transient Receptor Potential Channels; D012430:Ruthenium Red; D008536:Meglumine; C034516:dimethylamine; C004471:allyl isothiocyanate; C027917:tetramethylammonium; C023336:trimethylamine; D002118:Calcium