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. 2020 Aug 4;10(8):1142. doi: 10.3390/biom10081142

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Ca²⁺ binding to TgCEN1-C and TgCEN2-C. (A,B) Representative Ca²⁺ titration of apo-TgCEN1-C (A) and apo-TgCEN2-C (B) as monitored by ITC. Representative raw heat-power changes (upper panels) and integrated binding isotherms (bottom panels). A first injection of 0.5 μL was made and then the first data point was removed from data fitting. Curve fitting was performed by considering the two binding sites model for TgCEN1-C and the one site model for TgCEN2-C. The protein concentration was 70 μM and 100 μM, for apo-TgCEN1-C and apo-TgCEN2-C, respectively. (C) The downfield region of the ¹H-¹⁵N HSQC NMR spectra of TgCEN1-C recorded as a function of increasing Ca²⁺ concentration. The molar ratio of Ca²⁺:protein in each case was 0.9, 1.5, 4, and 10 (from left to right).