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Binding Interaction Between Prazosin and Immobilized Receptor by Frontal Analysis

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Abstract

Binding interaction between ligand and G-protein coupled receptor plays an important role in the several steps of exertion of therapeutic effect by a drug and has become increasingly interesting to pharmacists, chemists, biologists and companies. α1-Adrenoceptor (α1-AR) was purified from cell line which stably expressed α1-AR and evenly immobilized on the surface of macroporous silica gel to prepare a novel stationary phase for investigating the interaction between drug and receptor. Control drugs of α1-AR including phentolamine, terazosin and urapidil were used to characterize the retention properties of the stationary phase containing the receptor. Further work was performed to calculate the binding sites and association constant of prazosin binding to the immobilized receptor. The results presented a value of 4.55 × 10−7 M−1 for binding sites and of 2.2 × 104 M for the association constant during the interaction between prazosin and α1-AR. The proposed affinity method was stable at least in seven consecutive days, as well as had the primary bioactivities of recognizing and binding the ligand, providing an alternative for representing the interaction between drug and functional protein.

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Acknowledgments

Financial support was kindly provided from the National Natural Sciences Foundation of China (Nos. 21005060), the Chinese Ministry of Education (No. 20106101110001), Technology Support Plan Project (No. 2008BAI51B01) the Department of Education of the Shaanxi Province (No. 11JK0661), and the Key Innovative Project (2010ZDKG-46) of Science and Technology from Shaanxi Province.

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Authors and Affiliations

  1. College of Life Sciences, Northwest University, 195#, No. 229, Taibai North Road, Xi’an, 710069, Shaanxi, China

    Xinfeng Zhao, Haiyan Lu, Jing Huang & Xiaohui Zheng

  2. Institute of Analytical Science, Northwest University, Xi’an, 710069, China

    Jianbin Zheng

  3. Institute of Vascular Medicine, Peking University, Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences Ministry of Education, Beijing, 100083, China

    Youyi Zhang

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  1. Xinfeng Zhao
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  2. Haiyan Lu
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  3. Jing Huang
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  4. Jianbin Zheng
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  5. Xiaohui Zheng
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Correspondence to Xiaohui Zheng or Youyi Zhang.

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Zhao, X., Lu, H., Huang, J. et al. Binding Interaction Between Prazosin and Immobilized Receptor by Frontal Analysis. Chromatographia 75, 411–415 (2012). https://doi.org/10.1007/s10337-012-2198-4

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  • DOI: https://doi.org/10.1007/s10337-012-2198-4

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