Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 4 de 4
Filtrar
Mais filtros

Base de dados
Tipo de documento
País de afiliação
Intervalo de ano de publicação
1.
J Biomol Struct Dyn ; 36(8): 2147-2162, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28627969

RESUMO

Fasciola gigantica is the causative organism of fascioliasis and is responsible for major economic losses in livestock production globally. F. gigantica thioredoxin1 (FgTrx1) is an important redox-active enzyme involved in maintaining the redox homeostasis in the cell. To identify a potential anti-fasciolid compound, we conducted a structure-based virtual screening of natural compounds from the ZINC database (n = 1,67,740) against the FgTrx1 structure. The ligands were docked against FgTrx1 and 309 ligands were found to have better docking score. These compounds were evaluated for Lipinski and ADMET prediction, and 30 compounds were found to fit well for re-docking studies. After refinement by molecular docking and drug-likeness analysis, three potential inhibitors (ZINC15970091, ZINC9312362, and ZINC9312661) were identified. These three ligands were further subjected to molecular dynamics simulation (MDS) to compare the dynamics and stability of the protein structure after binding of the ligands. The binding free energy analyses were calculated to determine the intermolecular interactions. The results suggested that the two compounds had a binding free energy of -82.237, and -109.52 kJ.mol-1 for compounds with IDs ZINC9312362 and ZINC9312661, respectively. These predicted compounds displayed considerable pharmacological and structural properties to be drug candidates. We concluded that these two compounds could be potential drug candidates to fight against F. gigantica parasites.


Assuntos
Produtos Biológicos/química , Proteínas de Helminto/química , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Tiorredoxinas/química , Sequência de Aminoácidos , Animais , Produtos Biológicos/metabolismo , Produtos Biológicos/farmacologia , Avaliação Pré-Clínica de Medicamentos , Fasciola/genética , Fasciola/metabolismo , Proteínas de Helminto/antagonistas & inibidores , Proteínas de Helminto/metabolismo , Ligantes , Estrutura Molecular , Ligação Proteica , Domínios Proteicos , Homologia de Sequência de Aminoácidos , Termodinâmica , Tiorredoxinas/antagonistas & inibidores , Tiorredoxinas/metabolismo
2.
Exp Parasitol ; 151-152: 1-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25655406

RESUMO

A full-length complementary DNA (cDNA) encoding Cu/Zn-superoxide dismutase was isolated from Fasciola gigantica that on nucleotide sequencing showed a close homology (98.9%) with Cu/Zn-superoxide dismutase (SOD) of the temperate liver fluke, F. hepatica. Expression of the gene was found in all the three developmental stages of the parasite viz. adult, newly excysted juvenile and metacercaria at transcriptional level by reverse transcription-polymerase chain reaction (RT-PCR) and at the protein level by Western blotting. F. gigantica Cu/Zn-SOD cDNA was cloned and expressed in Escherichia coli. Enzyme activity of the recombinant protein was determined by nitroblue tetrazolium (NBT)-polyacrylamide gel electrophoresis (PAGE) and this activity was inactivated by hydrogen peroxide but not by sodium azide, indicating that the recombinant protein is Cu/Zn-SOD. The enzyme activity was relatively stable at a broad pH range of pH 4.0-10.0. Native Cu/Zn-superoxide dismutase protein was detected in the somatic extract and excretory-secretory products of the adult F. gigantica by Western blotting. NBT-PAGE showed a single Cu/Zn-SOD present in the somatic extract while three SODs are released ex vivo by the adult parasite. The recombinant superoxide dismutase did not react with the serum from buffaloes infected with F. gigantica. The role of this enzyme in defense by the parasite against the host reactive oxygen species is discussed.


Assuntos
DNA Complementar/isolamento & purificação , Fasciola/enzimologia , Regulação Enzimológica da Expressão Gênica , Superóxido Dismutase/isolamento & purificação , Matadouros , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Búfalos/parasitologia , DNA Complementar/química , DNA de Helmintos/química , DNA de Helmintos/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Fasciola/genética , Fasciola/crescimento & desenvolvimento , Fasciola hepatica/enzimologia , Fasciola hepatica/genética , Fasciolíase/parasitologia , Fasciolíase/veterinária , Concentração de Íons de Hidrogênio , Indicadores e Reagentes , Estágios do Ciclo de Vida/genética , Nitroazul de Tetrazólio , RNA de Helmintos/genética , RNA de Helmintos/isolamento & purificação , Coelhos , Proteínas Recombinantes/química , Análise de Sequência de DNA , Superóxido Dismutase/química , Superóxido Dismutase/genética
3.
Exp Parasitol ; 131(3): 283-91, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22543100

RESUMO

M17 leucine aminopeptidase (LAP) is one of a family of metalloexopeptidases, of which short peptide fragments are cleaved from the N-terminals. In this study, the full length of cDNA encoding Fasciola gigantica LAP (FgLAP) was cloned from adult parasites. The amino acid sequences of FgLAP showed a high degree of identity (98%) with that from Fasciola hepatica and a low degree of identities (11% and 9%) with those from cattle and human. Phylogenetic analysis revealed that the FgLAP was closely related and grouped with F. hepatica LAP (FhLAP). Northern analysis showed that FgLAP transcriptional products have 1800 base pairs. Analysis by RNA in situ hybridization indicated that LAP gene was expressed in the cecal epithelial cells of adult parasites. A polyclonal antibody to a recombinant FgLAP (rFgLAP) detected the native LAP protein in various developmental stages of the parasite. In a functional test, this rFgLAP displayed aminolytic activity using a fluorogenic Leu-MCA substrate, and was significantly inhibited by bestatin. Its maximum activity was at pH 8.0 and enhanced by Mn(2+) ions. Localization of LAP proteins by immunohistochemistry and immunofluorescence techniques indicated that the enzyme was distributed in the apical cytoplasm of cecal epithelial cells. Because of its important metabolic role and fairly exposed position, FgLAP is a potential drug target and a possible vaccine candidate against fasciolosis.


Assuntos
Clonagem Molecular , Fasciola/enzimologia , Leucil Aminopeptidase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Cricetinae , DNA Complementar/química , DNA de Helmintos/química , Fasciola/classificação , Fasciola/genética , Fasciola hepatica/classificação , Fasciola hepatica/enzimologia , Fasciola hepatica/genética , Feminino , Humanos , Leucil Aminopeptidase/química , Leucil Aminopeptidase/classificação , Lymnaea/parasitologia , Masculino , Mesocricetus , Dados de Sequência Molecular , Filogenia , RNA de Helmintos/análise , RNA Mensageiro/análise , Alinhamento de Sequência
4.
Mol Biochem Parasitol ; 146(1): 10-23, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16297461

RESUMO

A full-length cDNA encoding the Fasciola gigantica calcium-binding protein 1 (FgCaBP1) was cloned from an adult stage cDNA expression library in an immunoscreen using rabbit immune serum against the parasite's excretion/secretion antigens. The deduced amino acid sequence showed 96.3% identity to Fh22CBP of Fasciola hepatica. During development in the mammalian host FgCaBP1 RNA was detected in metacercariae, juveniles and adults and was exclusively localized to the tegumental cell bodies. Immune serum of a rabbit infected with F. gigantica detected recombinant FgCaBP1 starting from the sixth week of infection. Immune sera of mice infected with Schistosoma mansoni and Schistosoma mekongi cross-reacted with recombinant FgCaBP1 in immunoblots. Recombinant FgCaBP1 showed calcium and magnesium-binding activity by a mobility shift during non-denaturing PAGE in the presence of Ca2+ or Mg2+, respectively. A polyclonal mouse anti-rFgCaBP1 antiserum detected the native protein as a major component of the parasite's tegumental antigens in immunoblots and as a strictly tegumental antigen in tissue cross-sections of adult and juvenile parasites. Comparative sequence analysis of homologs from Fasciola and Schistosoma present in the GenBank database revealed sequence signatures specific to these trematode proteins and thereby indicates their origin from a single ancestor. FgCaBP1 contains two adjacent, N-terminal located EF-hands and a C-terminal located domain similar to dynein light chain type 1. Independent structure predictions of the two domains suggest that they will fold according to the already determined structures of the EF-hand motif and the dynein light chain type 1 proteins.


Assuntos
Proteínas de Ligação ao Cálcio/química , Proteínas de Ligação ao Cálcio/genética , Fasciola/metabolismo , Platelmintos/metabolismo , Sequência de Aminoácidos , Animais , Cálcio/farmacologia , Proteínas de Ligação ao Cálcio/imunologia , Proteínas de Ligação ao Cálcio/isolamento & purificação , Proteínas de Transporte/química , Bovinos , Clonagem Molecular/métodos , DNA Complementar/química , DNA Complementar/genética , Bases de Dados Genéticas , Proteínas de Drosophila/química , Dineínas , Motivos EF Hand/genética , Ensaio de Desvio de Mobilidade Eletroforética , Fasciola/genética , Fasciola/crescimento & desenvolvimento , Fasciola/imunologia , Fasciolíase/imunologia , Fasciolíase/parasitologia , Sequências Hélice-Alça-Hélice/genética , Camundongos , Camundongos Endogâmicos ICR , Dados de Sequência Molecular , Filogenia , Platelmintos/classificação , Platelmintos/genética , Coelhos , Proteínas Recombinantes/isolamento & purificação , Alinhamento de Sequência , Análise de Sequência de Proteína
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA