| Full text | |
| 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: | 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 Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 07/08703-0 - Design of inhibitors for dihydroorotate dehydrogenase from Trypanosoma cruzi and Leishmania major |
| Grantee: | Matheus Pinto Pinheiro |
| Support Opportunities: | 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 Opportunities: | Scholarships in Brazil - Doctorate (Direct) |
| FAPESP's process: | 11/14269-6 - Crystallographic studies of Dihydroorotate dehydrogenase from schistosoma mansoni |
| Grantee: | Giovani Pinton Tomaleri |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |
| 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 Opportunities: | Regular Research Grants |