| Author | Gadelha, Ana Paula R. | |
| Author | Brigagao, Claudia Maia | |
| Author | Silva, Martha Barros da | |
| Author | Rodrigues, Aline Beatriz Mello | |
| Author | Guimarães, Ana Carolina Ramos | |
| Author | Paiva, Fernando | |
| Author | Souza, Wanderley de | |
| Author | Henriques, Cristina | |
| Access date | 2020-06-01T14:37:39Z | |
| Available date | 2020-06-01T14:37:39Z | |
| Document date | 2020 | |
| Citation | GADELHA, Ana Paula R. et al. Insights about the structure of farnesyl diphosphate synthase (FPPS) and the activity of bisphosphonates on the proliferation and ultrastructure of Leishmania and Giardia. Parasites & Vectors, v. 13, n. 168, p. 1-18, 2020. | pt_BR |
| ISSN | 1756-3305 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/41468 | |
| Language | eng | pt_BR |
| Publisher | BMC | pt_BR |
| Rights | open access | |
| Subject in Portuguese | Biofosfonatos | pt_BR |
| Subject in Portuguese | Sinsfostato de difosfato farnezyl | pt_BR |
| Subject in Portuguese | Leishmania | pt_BR |
| Subject in Portuguese | Protozoário | pt_BR |
| Subject in Portuguese | Esterol | pt_BR |
| Subject in Portuguese | Ergosterol | pt_BR |
| Title | Insights about the structure of farnesyl diphosphate synthase (FPPS) and the activity of bisphosphonates on the proliferation and ultrastructure of Leishmania and Giardia | pt_BR |
| Type | Article | |
| DOI | 10.1186/s13071-020-04019-z | pt_BR |
| Abstract | Background: The enzyme farnesyl diphosphate synthase (FPPS) is positioned in the intersection of diferent sterol biosynthesis pathways such as those producing isoprenoids, dolichols and ergosterol. FPPS is ubiquitous in eukaryotes and is inhibited by nitrogen-containing bisphosphonates (N-BP). N-BP activity and the mechanisms of cell death as well as damage to the ultrastructure due to N-BP has not yet been investigated in Leishmania infantum and Giardia. Thus, we evaluated the efect of N-BP on cell viability and ultrastructure and then performed structural modelling and phylogenetic analysis on the FPPS enzymes of Leishmania and Giardia. Methods: We performed multiple sequence alignment with MAFFT, phylogenetic analysis with MEGA7, and 3D structural modelling for FPPS with Modeller 9.18 and on I-Tasser server. We performed concentration curves with N-BP in Leishmania promastigotes and Giardia trophozoites to estimate the IC50 via the MTS/PMS viability method. The ultrastructure was evaluated by transmission electron microscopy, and the mechanism of cell death by fow cytometry. Results: The nitrogen-containing bisphosphonate risedronate had stronger anti-proliferative activity in Leishmania compared to other N-BPs with an IC50 of 13.8 µM, followed by ibandronate and alendronate with IC50 values of 85.1 µM and 112.2 µM, respectively. The efect of N-BPs was much lower on trophozoites of Giardia than Leishmania (IC50 of 311 µM for risedronate). Giardia treated with N-BP displayed concentric membranes around the nucleus and nuclear pyknosis. Leishmania had mitochondrial swelling, myelin fgures, double membranes, and plasma membrane blebbing. The same population labelled with annexin-V and 7-AAD had a loss of membrane potential (TMRE), indicative of apoptosis. Multiple sequence alignments and structural alignments of FPPS proteins showed that Giardia and Leishmania FPPS display low amino acid identity but possess the conserved aspartate-rich motifs. Conclusions: Giardia and Leishmania FPPS enzymes are phylogenetically distant but display conserved protein signatures. The N-BPs efect on FPPS was more pronounced in Leishmania than Giardia. This might be due to general diferences in metabolism and diferences in the FPPS catalytic site. | pt_BR |
| Affilliation | Instituto Nacional de Metrologia. Laboratório de Biotecnologia. Rio de Janeiro, RJ, Brasil / Universidade do Grande Rio. Duque de Caxias, RJ, Brasil / Universidade Federal do Rio de Janeiro Laboratório de Ultraestrutura Celular Herta Meyer. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro. Laboratório de Ultraestrutura Celular. Hertha Meyer. Rio de Janeiro, RJ, Brasil. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Mato Grosso do Sul. Centro de Ciências Biológicas e da Saúde. Departamento de Patologia. Campo Grande, MS, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Fiocruz Mato Grosso do Sul. Campo Grande, MS, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biotecnologia e Fisiologia de Infecções Virais. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | Biophosphonates | pt_BR |
| Subject | Farnesyl diphosphate synthase | pt_BR |
| Subject | FPPS | pt_BR |
| Subject | Leishmania | pt_BR |
| Subject | Giardia | pt_BR |
| Subject | Protozoan | pt_BR |
| Subject | Isoprenoid | pt_BR |
| Subject | Isoprenylation | pt_BR |
| Subject | Sterol | pt_BR |
| Subject | Ergosterol | pt_BR |
