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. 2019 Dec 9;9:18629. doi: 10.1038/s41598-019-54970-w

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© The Author(s) 2019

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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SPR sensorgrams (black curves) of the response (Resonance Units, RU) versus time of the single-cycle kinetics performed by injecting five increasing concentrations (0.01, 0.05, 0.1, 0.5, 1 µM) of Hb over the IsdB-functionalized substrate. The red curves are the fitting with either a 1:1 or a two state irreversible binding model. Fitting parameters are reported in Table 2. The resulting kinetic parameters are reported in Table 2. (A) IsdB-OxyHb (1:1) (B) IsdB-α-semiOxyHb (1:1) (C) IsdB-β-semiOxyHb (1:1) (D) IsdB-MetHb (two-state irreversible) (E) IsdB-α-semiMetHb (two-state irreversible) (F) IsdB-β-semiMetHb (two-state irreversible) (G) IsdB^Y440A-OxyHb (1:1) (H) IsdB^Y440A-MetHb (two-state irreversible).