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. 2019 Jan 22;9:300. doi: 10.1038/s41598-018-36391-3

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© The Author(s) 2019

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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ITC binding measurements. Binding of 2-isobutyl-3-methoxypyrazine (PYR) and menadione (MEN) to wild type and mutant MUPs. ITC data for PYR (A) and MEN (B) binding to recombinant wild type MUP (WT) (a), and mutants L88Q (b), L124V (c) and Y103R (d). Raw injection heats (expressed as differential power) are shown in the top panels and the corresponding specific binding isotherms, calculated from the integrated injection heats and normalized to the moles of injectant, are shown in the bottom panels. The equilibrium dissociation constants (K_d) and stoichiometry values (N) were determined by fitting the data to a one-site binding model.