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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Purification and characterization of a specific late-larval esterase from two species of the Drosophila repleta group: contributions to understand its evolution

Texto completo
Lopes, Vanessa F. [1] ; Cabral, Hamilton [2] ; Machado, Luciana P. B. [1] ; Mateus, Rogerio P. [1]
Número total de Autores: 4
Afiliação do(s) autor(es):
[1] Univ Estadual Ctr Oeste UNICENTRO, Dept Ciencias Biol, Lab Genet & Evolucao, BR-85040080 Guarapuava, Parana - Brazil
[2] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Lab Tecnol Enzimat, BR-14040903 Ribeirao Preto, SP - Brazil
Número total de Afiliações: 2
Tipo de documento: Artigo Científico
Fonte: ZOOLOGICAL STUDIES; v. 53, JAN 2014.
Citações Web of Science: 2

Background: After duplication, one copy of an original gene can become redundant and decay toward a pseudogene status or functionally diverge. Here, we performed the purification and biochemical characterization of EST-4 (a late larval beta-esterase) from two Drosophila repleta group species, Drosophila mulleri and Drosophila arizonae, in order to establish comparative parameters between these enzymes in these species and to contribute to better understand their evolution. Results: In D. mulleri, EST-4 had an optimal activity in temperatures ranging from 40 degrees to 45 degrees C and at pH 7.5, maintaining stability in alkaline pH (8.0 to 10.0). It was classified as serine esterase as its activity was inhibited by PMSF. No ion negatively modulated EST-4 activity, and iron had the most positive modulating effect. In D. arizonae, it showed similar optimum temperature (40 degrees C), pH (8.0), and was also classified as a serine esterase, but the enzymatic stability was maintained in an acidic pH (5.5 to 6.5). Fe+2 had the opposite effect found in D. mulleri, that is, negative modulation. Al+3 almost totally inhibited the EST-4 activity, and Na+ and Cu+2 had a positive modulation effect. Kinetic studies, using rho-nitrophenyl acetate as substrate, showed that EST-4 from D. mulleri had higher affinity, while in D. arizonae, it showed higher V-max and catalytic efficiency in optimal reaction conditions. Conclusions: EST-4 from D. mulleri and D. arizonae are very closely related and still maintain several similar features; however, they show some degree of differentiation. Considering that EST-4 from D. mulleri has more conspicuous gel mobility difference among all EST-4 studied so far and a lower catalytic efficiency was observed here, we proposed that after duplication, this new copy of the original gene became redundant and started to decay toward a pseudogene status in this species, which probably is not occurring in D. arizonae. (AU)

Processo FAPESP: 11/06986-0 - Determinação da especificidade de peptidases isoladas de fungos usando peptídeos FRET como substratos
Beneficiário:Hamilton Cabral
Linha de fomento: Auxílio à Pesquisa - Regular