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. 2017 Jun 2;7:2701. doi: 10.1038/s41598-017-02953-0

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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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Maximum likelihood tree of the aminergic toxins. The best evolutionary model established by MEGA5 was the Dayhoff model using a discrete Gamma distribution (+G). Bootstraps value higher than 50% are shown. MTLP-1 and 2 were used as out-group. Six ancestral sequences are figured on the nodes of the aminergic toxins tree. Quality of the reconstruction is shown on right side upper panel. For each site, a probability P is given. In total 390 sites were reconstructed. Among them, only 31 showed a P < 0.9, highlighting a good quality of the reconstruction. As shown in the lower panel, a high proportion of these ambiguous sites are presented in the sequences of AncTx1 and 2. The ambiguities were resolved by chosen the residues with the best probability. Due to the high divergence of MT7 and MT2 in term of sequence and function, this choice leads AncTx1 and 2 to be closer of AncTx3 and ρ-Da1a than MT7/MT2.