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. 2018 May 18;8:7872. doi: 10.1038/s41598-018-26213-x

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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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Stopped-flow kinetics for the binding between TAZ1 and Hif-1α CAD. (A) Example of a binding kinetics trace for TAZ1/Hif-1α CAD (0.5 μM/8 μM), and a double-exponential function was used for fitting. The inset shows a zoomed view of the fast phase. (B) The concentration dependence of k_obs for the slow (inset) and fast phase. The k_obs for the fast phase is linearly dependent on Hif-1α CAD concentration while the k_obs for the slow phase is concentration independent, with a value of 3–4 s⁻¹ for all Hif-1α CAD variants. k_on^app was obtained by fitting the concentration dependence of k_obs for the fast phase to the general equation for association of two molecules²⁸.