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Sci. Lett. J. 2012, 1: 25
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Research Article
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Full Text
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Determination of the binding parameters between lysozyme and dihydroxybenzenes by flow injection chemiluminescence analysis
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Minxia Shen, Xijuan Tan, Haixiang Zhao, Zhenghua Song
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Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, College of Chemistry and Materials Science, Northwest University, Xi’an, 710069, China
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Abstract |
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Based on the quenching effect of dihydroxybenzenes (hydroquinone HQ, catechol CT and resorcinol RS) on luminol–lysozyme chemiluminescence (CL) system, the binding parameters of lysozyme with dihydroxybenzenes were first determined by flow injection chemiluminescence (FI-CL) analysis. It was found that the CL intensity decrements were logarithm over the concentrations of dihydroxybenzenes, giving a general equation of ΔI = ALnC + B (R > 0.99). At a flow rate of 2.0 mL min–1, a complete analysis, including sampling and washing, could be performed in 25 s with relative standard deviations (RSDs) of less than 3.0% (n = 5). By homemade FI-CL model of protein–small molecule interaction, lg[(I0 – I)/I] = lgKD + nlg[D], the binding constants KD (at 105–107 level) and the number of interacting sites n (approximately 1) were obtained; the affinity of dihydroxybenzenes to lysozyme followed the order: HQ > CT > RS; and the reaction should be spontaneous with hydrophobic effect.
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Keywords
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Lysozyme; Dihydroxybenzenes; Binding parameters; Chemiluminescence; Flow injection analysis
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