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. 2017 Sep 8;7:10999. doi: 10.1038/s41598-017-11194-0

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Effects of individual TRPM8 point mutations on calcium responses induced by cooling agent 10 and icilin. Shown data represent mean ± standard error of the mean (SEM) of quadruplicate determinations of a representative experiment (n = 4). Statistical analysis was performed using unpaired Student’s t test with GraphPad Prism (95% confidence interval). (A) Dose response curves of icilin on mutant TRPM8 receptors. The substitutions Y745A, D802A and N799A abrogated agonist activity. The I746A substitution significantly (p < 0.05) shifted pIC₅₀ of Icilin from 6.6 to 6.9. (B) Dose response curves of Cooling Agent 10 on mutant TRPM8 receptors. The substitution I746A strongly reduced agonist activity in term of efficacy while D802A significantly (p < 0.0001) shifted the pIC₅₀ of the agonist from 5.1 to 4.5. Other mutations did not have effects on Cooling agent 10 activity.