| Full text | |
| Author(s): |
Calil, Felipe Antunes
;
Lima, Juliana Maria
;
Cavalcante de Oliveira, Arthur Henrique
;
Mariotini-Moura, Christiane
;
Rangel Fietto, Juliana Lopes
;
Cardoso, Carmen Lucia
Total Authors: 6
|
| Document type: | Journal article |
| Source: | JOURNAL OF ANALYTICAL METHODS IN CHEMISTRY; v. 2016, p. 9-pg., 2016-01-01. |
| Abstract | |
The use of IMERs (Immobilized Enzyme Reactors) as a stationary phase coupled to high performance chromatographic systems is an interesting approach in the screening of new ligands. In addition, IMERs offer many advantages over techniques that employ enzymes in solution. The enzyme nucleoside triphosphate diphosphohydrolase (NTPDase-1) from Trypanosoma cruzi acts as a pathogen infection facilitator, so it is a good target in the search for inhibitors. In this paper, immobilization of NTPDase-1 afforded ICERs (Immobilized Capillary Enzyme Reactors). A liquid chromatography method was developed and validated to monitor the ICER activity. The conditions for the application of these bioreactors were investigated, and excellent results were obtained. The enzyme was successfully immobilized, as attested by the catalytic activity detected in the TcNTPDase-1-ICER chromatographic system. Kinetic studies on the substrate ATP gave K-M of 0.317 +/- 0.044 mmol.L-1, which still presented high affinity compared to in solution. Besides that, the ICER was stable for 32 days, enough time to investigate samples of possible inhibitors, including especially the compound Suramin, that inhibited 51% the enzyme activity at 100 mu mol.L-1, which is in accordance with the data for the enzyme in solution. (AU) | |
| FAPESP's process: | 13/01710-1 - Enzyme ligand: new models of screening |
| Grantee: | Quezia Bezerra Cass |
| Support Opportunities: | Research Projects - Thematic Grants |
| FAPESP's process: | 08/04371-5 - Development and applications of enzymatic columns in selective inhibitors screening and in the natural products studies |
| Grantee: | Carmen Lúcia Cardoso |
| Support Opportunities: | Research Grants - Young Investigators Grants |