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. 2017 Dec 20;7:17933. doi: 10.1038/s41598-017-18085-4

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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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Isothermal titration microcalorimetric analysis of Ca²⁺ and Zn²⁺ binding to FSA (a,c) and EH-CC-E204G (b,d). Trace of the calorimetric titration of 20 × 2-μl aliquots of 15 mM CaCl₂ or 30 (36) × 1-μl aliquots of 15 mM ZnCl₂ into 50 μM proteins (top), and integrated binding isotherms (bottom).