TRP channels sense temperatures ranging from noxious cold to noxious heat. Whether specialized TRP thermosensor modules exist and how they control channel pore gating is unknown. We studied purified human TRPA1 (hTRPA1) truncated proteins to gain insight into the temperature gating of hTRPA1. In patch-clamp bilayer recordings, ∆1-688 hTRPA1, without the N-terminal ankyrin repeat domain (N-ARD), was more sensitive to cold and heat, whereas ∆1-854 hTRPA1, also lacking the S1-S4 voltage sensing-like domain (VSLD), gained sensitivity to cold but lost its heat sensitivity. In hTRPA1 intrinsic tryptophan fluorescence studies, cold and heat evoked rearrangement of VSLD and the C-terminus domain distal to the transmembrane pore domain S5-S6 (CTD). In whole-cell electrophysiology experiments, replacement of the CTD located cysteines 1021 and 1025 with alanine modulated hTRPA1 cold responses. It is proposed that hTRPA1 CTD harbors cold and heat sensitive domains allosterically coupled to the S5-S6 pore region and the VSLD, respectively.

Department of Biomedical and Clinical Sciences Faculty of Health Sciences Linköping University SE 581 83 Linköping Sweden

Department of Cellular Neurophysiology Institute of Physiology of the Czech Academy of Sciences 142 20 Prague Czech Republic

Department of Clinical Sciences Malmö Lund University SE 214 28 Malmö Sweden

Department of Physics Chemistry and Biology Division of Chemistry Linköping University SE 58183 Linköping Sweden

Janssen Research and Development Division of Janssen Pharmaceutica N 5 Turnhoutseweg 30 2340 Beerse Belgium

Leibniz Institut für Analytische Wissenschaften ISAS e 5 Bunsen Kirchhoff Straße 11 44139 Dortmund Germany

Wallenberg Centre for Molecular Medicine Linköping University SE 581 83 Linköping Sweden

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Human Transient Receptor Potential Ankyrin 1 Channel: Structure, Function, and Physiology