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. 2020 Dec 4;10:21309. doi: 10.1038/s41598-020-78269-3

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Characterization of labeling sites on CarD. (A) Sites on CarD which were selected for site-specific mutagenesis and subsequent labeling with BODIPY-FL (colors correspond with binding isotherms in (B). (B) Binding isotherms for CarD mutants labeled with BODIPY-FL (note: several differences in this assay are as follows: 50 mM KGlu. 0.001% Triton X-100, and 12.5 nM CarD-BODIPY-FL were used in this experiment). (C) Summary of isotherm parameters for each CarD variant as well as domain interaction on the RPo where the BODIPY-FL probe is located.