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. 2021 Mar 1;12:1347. doi: 10.1038/s41467-021-21548-y

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© The Author(s) 2021

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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Fig. 2 — a Relative activity of single substitutions, UrdA′_R411A, UrdA′_R560A, and UrdA′_D388A compared to UrdA′_wt (n = 6 independent experiments). Data are mean ± s.e.m. ***P < 0.001. One-way ANOVA with Dunnett’s multiple comparisons test, P value < 0.0001. b Activity and specificity of the full-length UrdA toward varying concentrations of urocanate (n = 3 independent experiments) or fumarate (n = 3 independent experiments) measured in presence of FAD. c Relative activity of single substitution variants, UrdA_R411A, UrdA_R560A, and UrdA_D388A compared to the full-length UrdA_wt in an absorbance-based measurement (left, n = 3 independent experiments) and in a direct measurement of ImP production (right, n = 3 independent experiments). Data are mean ± s.e.m. ***P < 0.001. One-way ANOVA with Dunnett’s multiple comparisons test, P value < 0.0001. d ITC measurement of urocanate (blue) and of ImP (orange) binding to UrdA′, showing a representative dataset for each ligand. The full line shows the simultaneous fit to three replicate datasets. Errors in individual ITC data points are estimated based on the baseline uncertainty, as described³⁶. Source data are provided as a Source data file.