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Determining PPARγ-ligand binding affinity using fluorescent assay with cis-parinaric acid as a probe

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Abstract

Upon the study of small-molecules binding to proteins, the traditional methods for calculating dissociation constants (K d and K i ) have shortcomings in dealing with the single binding site models. In this paper, two equations have been derived to solve this problem. These two equations are independent of the total concentration or initial degree of saturation of receptor and the activity of the competitive molecule. Through nonlinear fitting against these two equations, K d value of a probe can be obtained by binding assay, and K i value of a ligand can be obtained by competitive assay. Moreover, only the total concentrations of receptor([R]t), ligand([L]t) and probe([P]t) are required for the data fitting. In this work, K i values of some typical ligands of PPARγ were successfully determined by use of our equations, among which the K i value of PPARγ-LY171883 was reported for the first time.

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Authors and Affiliations

  1. Drug Discovery and Design Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 201203, Shanghai, China

    Zhenting Gao, Haibin Luo, Lili Chen, Jianhua Shen, Kaixian Chen, Hualiang Jiang & Xu Shen

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  1. Zhenting Gao
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  2. Haibin Luo
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  3. Lili Chen
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  4. Jianhua Shen
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  5. Kaixian Chen
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  6. Hualiang Jiang
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  7. Xu Shen
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Correspondence to Jianhua Shen or Hualiang Jiang.

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Gao, Z., Luo, H., Chen, L. et al. Determining PPARγ-ligand binding affinity using fluorescent assay with cis-parinaric acid as a probe. Sc. China Ser. B-Chem. 48, 122–131 (2005). https://doi.org/10.1360/03yb0217

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  • DOI: https://doi.org/10.1360/03yb0217

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