Nature Neuroscience 9, 493 - 500 (2006)
Published online: 5 March 2006; Corrected online: 19 March 2006 | doi:10.1038/nn1665
High-throughput random mutagenesis screen reveals TRPM8 residues specifically required for activation by mentholMichael Bandell1, 2, Adrienne E Dubin3, Matt J Petrus2, Anthony Orth2, Jayanti Mathur2, Sun Wook Hwang1, 4
& Ardem Patapoutian1, 21
Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA. 2
Genomics Institute of the Novartis Research Foundation, San Diego, California 92121, USA. 3
Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA. 4
Korea University Graduate School of Medicine, Seoul 136-705, Korea
Correspondence should be addressed to Ardem Patapoutian ardem@scripps.edu Menthol is a cooling compound derived from mint leaves and is extensively used as a flavoring chemical. Menthol activates transient receptor potential melastatin 8 (TRPM8), an ion channel also activated by cold, voltage and phosphatidylinositol-4,5-bisphosphate (PIP2). Here we investigated the mechanism by which menthol activates mouse TRPM8. Using a new high-throughput approach, we screened a random mutant library consisting of  14,000 individual TRPM8 mutants for clones that are affected in their response to menthol while retaining channel function. We identified determinants of menthol sensitivity in two regions: putative transmembrane segment 2 (S2) and the C-terminal TRP domain. Analysis of these mutants indicated that activation by menthol involves a gating mechanism distinct and separable from gating by cold, voltage or PIP2. Notably, TRP domain mutations mainly attenuated menthol efficacy, suggesting that this domain influences events downstream of initial binding. In contrast, S2 mutations strongly shifted the concentration dependence of menthol activation, raising the possibility that S2 influences menthol binding.Note: The AOP version of this article was corrected on 19 March 2006. Please see the PDF for details.
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