| Full text | |
| Author(s): |
Total Authors: 4
|
| Affiliation: | [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
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | ZOOLOGICAL STUDIES; v. 53, JAN 2014. |
| Web of Science Citations: | 2 |
| Abstract | |
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) | |
| FAPESP's process: | 11/06986-0 - Determination of the specificity of peptidases isolated from fungi using fluorescence resonance energy transfer (FRET) peptides as substrates |
| Grantee: | Hamilton Cabral |
| Support Opportunities: | Regular Research Grants |