Development of the Fluorescent Biosensor hCalmodulin (hCaM)L39C-monobromobimane(mBBr)/V91C-mBBr, a Novel Tool for Discovering New Calmodulin Inhibitors and Detecting

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• 作者：Marti虂n Gonza虂lez-Andrade ; Jose虂 Rivera-Cha虂vez ; Alejandro Sosa-Peinado ; Mario Figueroa ; Rogelio Rodri虂guez-Sotres ; Rachel Mata
• 刊名：Journal of Medicinal Chemistry
• 出版年：2011
• 出版时间：June 9, 2011
• 年：2011
• 卷：54
• 期：11
• 页码：3875-3884
• 全文大小：1145K
• 年卷期：v.54,no.11(June 9, 2011)
• ISSN：1520-4804

文摘

A novel, sensible, and specific fluorescent biosensor of human calmodulin (hCaM), namely hCaM L39C-mBBr/V91C-mBBr, was constructed. The biosensor was useful for detecting ligands with opposing fluorescent signals, calcium ions (Ca²⁺) and CaM inhibitors in solution. Thus, the device was successfully applied to analyze the allosteric effect of Ca²⁺ on trifluoroperazine (TFP) binding to CaM (Ca²⁺K_d = 0.24 渭M 卤 0.03 with a stoichiometry 4.10 卤 0.15; TFPK_d 5.74鈥?.53 渭M depending on the degree of saturation of Ca²⁺, with a stoichiometry of 2:1). In addition, it was suitable for discovering additional xanthones (5, 6, and 8) with anti-CaM properties from the fungus Emericella 25379. The affinity of 1鈥?b>5, 7, and 8 for the complex (Ca²⁺)₄-CaM was excellent because their experimental K_ds were in the nM range (4鈥?98 nM). Docking analysis predicted that 1鈥?b>8 bind to CaM at sites I, III, and IV as does TFP.