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. 2018 Apr 27;8:6635. doi: 10.1038/s41598-018-24493-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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Characterization of DCP-NEt₂C and DCP-Rho1 fluorescence and kinetic properties. (a) Chemical structure of DCP-Rho1. (b) DCP-NEt₂C synthesis scheme and structure. (c) Excitation and emission spectra of DCP-NEt₂C. Dotted line shows excitation spectrum taken with emission at λ_max 499 nm; solid line line shows emission spectrum taken with excitation at λ_max 421 nm. (d) Analysis of DCP-NEt₂C (blue) and DCP-Rho1 (red) dependence of fluorescence signal on pH. (e) Deconvoluted mass spectra of oxidized C165A AhpC (black), and its 60 min reaction with dimedone (green), DCP-NEt₂C (blue), and DCP-Rho1 (red). (f) SDS-PAGE fluorescence imaging analysis of oxidized C165A AhpC reaction with DCP-NEt₂C and DCP-Rho1. (g) Kinetic profiles and reaction rates of oxidized C165A AhpC reaction with dimedone (black circle), DCP-NEt₂C (open circle), and DCP-Rho1 (open square). The data were fit using single exponential kinetics and the second order rate constants were calculated by dividing the rate with the respective probe concentration.