ThermoFMN - a thermofluor assay developed for ligand-screening as an alternative strategy for drug discovery

Journal of the Brazilian Chemical Society

Ricardo A. P. Pádua¹  Giovani P. Tomaleri¹  Renata A. G. Reis¹  Juliana S. David¹  Valeria C. Silva¹  Matheus P. Pinheiro¹  Maria Cristina Nonato¹ 

[1] ,Universidade de São Paulo Faculdade de Ciências Farmacêuticas de Ribeirão Preto Laboratório de Cristalografia de ProteínasRibeirão Preto SP ,Brazil

关键词: FMN;    Thermofluor;    drug targets;    ThermoFMN;    drug discovery;    dihydroorotate dehydrogenase;   

DOI  :  10.5935/0103-5053.20140157

来源: SciELO

【 摘 要 】

Thermofluor has become a well-known and widely practiced methodology for screening of ligands that enhance stability and solubility of proteins, and also a powerful tool for hit identification in early drug discovery. In the present work, we developed an alternative Thermoflour method, named ThermoFMN, which explores the endogenous prosthetic group flavin mononucleotide (FMN) of flavoproteins as the fluorescent probe. Validation of ThermoFMN method was achieved by monitoring fluorescence signal of FMN of several drug targets in the presence of an unbiased library of ligands. In addition, drugs with known efficacy had their selective inhibition profile evaluated. Besides demonstrating that FMN signal provides sufficient fluorescence intensity for detection, our results revealed that ThermoFMN assay requires low concentration of protein samples and is compatible with a wide range of chemical reagents. The methodology presented here proposes an alternative strategy in the search for ligands of FMN-binding drug targets, therefore an important tool for the development of new therapies against neglected diseases.

【 授权许可】

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