Structure and kinetics assays of recombinant Schistosoma mansoni dihydrofolate reductase

Autor(es)

Balasco Serrao, Vitor Hugo ; Romanello, Larissa ; Cassago, Alexandre ; Torini de Souza, Juliana Roberta ; Cheleski, Juliana ; DeMarco, Ricardo ; Brandao-Neto, Jose ; Pereira, Humberto D'Muniz

• Tipo de documento: Artigo Científico
• Fonte: Acta Tropica; v. 170, p. 190-196, JUN 2017.
• Citações Web of Science: 3

Resumo

The parasite Schistosoma mansoni possesses all pathways for pyrimidine biosynthesis, in which dihydrofolate reductase (DHFR), thymidylate cycle participants, is essential for nucleotide metabolism to obtain energy and structural nucleic acids. Thus, DHFRs have been widely suggested as therapeutic targets for the treatment of infectious diseases. In this study, we expressed recombinant SmDHFR in a heterologous manner to obtain structural, biochemical and kinetic information. X-ray diffraction of recombinant SmDHFR at 1.95 angstrom resolution showed that the structure exhibited the canonical DHFR fold. Isothermal titration calorimetry was used to determine the kinetic constants for NADP(+) and dihydrofolate. Moreover, inhibition assays were performed using the commercial folate analogs methotrexate and aminopterin; these analogs are recognized as folate competitors and are used as chemotherapeutic agents in cancer and autoimmune diseases. This study provides information that may prove useful for the future discovery of novel drugs and for understanding these metabolic steps from this pathway of S. mansoni, thus aiding in our understanding of the function of these essential pathways for parasite metabolism. (C) 2017 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 12/14223-9 - Estudos estruturais e cinéticos das enzimas da via de salvação de purina de Schistosoma mansoni
• Beneficiário: Humberto D'Muniz Pereira
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 12/23730-1 - Caracterização das interações macromoleculares das proteínas envolvidas na síntese de selenocisteínas em escherichia coli
• Beneficiário: Vitor Hugo Balasco Serrão
• Modalidade de apoio: Bolsas no Brasil - Doutorado