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. 2018 Oct 4;8:14837. doi: 10.1038/s41598-018-33096-5

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© The Author(s) 2018

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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CHP3 binds with high affinity to the CBD region of NHE1 as probed in vitro by microscale thermophoresis. (a) Representative MST data for CHP3:MBP-CBD interaction showing superimposed time traces of five independent titration series for one biological replicate. The fluorescence was normalized to a starting value of 1. The laser was switched off after 5 s and the thermophoresis + T-jump signals were used for data analysis. Time spans for F_c and F_h are indicated with dotted and dashed lines, respectively. (b) MST binding curve derived from (a) as described in Materials and Methods. The line shows the best fit using the bimolecular binding model. The data for three biological replicates are summarized in Table 1. A mean K_D = 55.5 ± 6.8 nM (mean ± standard deviation (SD)) was determined.