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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

ThermoFMN - A Thermofluor Assay Developed for Ligand-Screening as an Alternative Strategy for Drug Discovery

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Author(s):
Padua, Ricardo A. P. [1] ; Tomaleri, Giovani P. [1] ; Reis, Renata A. G. [1] ; David, Juliana S. [1] ; Silva, Valeria C. [1] ; Pinheiro, Matheus P. [1] ; Nonato, Maria Cristina [1]
Total Authors: 7
Affiliation:
[1] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Lab Cristalog Proteinas, BR-14040903 Ribeirao Preto, SP - Brazil
Total Affiliations: 1
Document type: Journal article
Source: Journal of the Brazilian Chemical Society; v. 25, n. 10, p. 1864+, OCT 2014.
Web of Science Citations: 7
Abstract

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. (AU)

FAPESP's process: 07/08703-0 - Design of inhibitors for dihydroorotate dehydrogenase from Trypanosoma cruzi and Leishmania major
Grantee:Matheus Pinto Pinheiro
Support type: Scholarships in Brazil - Doctorate
FAPESP's process: 08/11644-8 - Structural and functional characterization of Trypanosoma cruzi fumarate hydratase isoforms
Grantee:Ricardo Augusto Pereira de Pádua
Support type: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 12/25075-0 - Development of leishmanicidal drugs based on the selective inhibition of the enzyme dihydroorotate dehydrogenase
Grantee:Maria Cristina Nonato
Support type: Regular Research Grants
FAPESP's process: 11/14269-6 - Crystallographic studies of Dihydroorotate dehydrogenase from schistosoma mansoni
Grantee:Giovani Pinton Tomaleri
Support type: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 11/23504-9 - Exploring structure-function relationship of DHODH enzyme in the development of new molecules with trypanocidal and leishmanicidal activities
Grantee:Renata Almeida Garcia Reis
Support type: Scholarships in Brazil - Doctorate