Abstract
TRPV1 is the founding and best-studied member of the family of temperature-activated transient receptor potential ion channels (thermoTRPs). Voltage, chemicals and heat allosterically gate TRPV1. Molecular determinants of TRPV1 activation by capsaicin, allicin, acid, ammonia and voltage have been identified. However, the structures and mechanisms mediating TRPV1's pronounced temperature sensitivity remain unclear. Recent studies of the related channel TRPV3 identified residues in the pore region that are required for heat activation. We used both random and targeted mutagenesis screens of rat TRPV1 and identified point mutations in the outer pore region that specifically impair temperature activation. Single-channel analysis indicated that TRPV1 mutations disrupted heat sensitivity by ablating long channel openings, which are part of the temperature-gating pathway. We propose that sequential occupancy of short and long open states on activation provides a mechanism for enhancing temperature sensitivity. Our results suggest that the outer pore is important for the heat sensitivity of thermoTRPs.
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Acknowledgements
We thank A. Marelli and T. Orth for preparing mini-prep DNA, M. Caterina for providing rat TRPV1 plasmid DNA, and B. Coste for critical reading of the manuscript. This research was supported by the US National Institutes of Health, the Novartis Research Foundation and a fellowship to J.G. from the American Heart Association.
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J.G. designed the study, collected and analyzed data, and wrote the manuscript. S.E.K. collected and analyzed data in the primary screen. V.U. prepared the mutant library and collected and analyzed data in the primary screen. B.B. performed molecular modeling. M.P. prepared the mutant library. M.B. developed the screening conditions. A.P. designed the study and edited the manuscript. All of the authors discussed the results and commented on the manuscript.
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Grandl, J., Kim, S., Uzzell, V. et al. Temperature-induced opening of TRPV1 ion channel is stabilized by the pore domain. Nat Neurosci 13, 708–714 (2010). https://doi.org/10.1038/nn.2552
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DOI: https://doi.org/10.1038/nn.2552
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