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. 2022 Feb 17;12:2691. doi: 10.1038/s41598-022-06591-z

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Assessment of the functional role of tunnels. (a) Mutation of A290^7.32 to W290^7.32 could lead to high steric hinderance in Tunnel 1 (green). (b) Mutation of L35^1.35 to W35^1.35 could lead to high steric hinderance in Tunnel 2 (yellow). (c) Dose-dependent response curves of wild type (WT), A290^7.32W single mutant, L35^1.35W single mutant, and A290^7.32W & L35^1.35W double mutant TAAR9 to spermidine. The A290^7.32W or L35^1.35W single mutant shows much lower maximal response and larger EC₅₀. The A290^7.32W & L35^1.35W double mutant shows further decrease in maximal response.