Purificação, caracterização físico-química e bioatividade de uma proteína inativadora de ribossomos de Jatropha podagrica Hook
Ribosome inactivating proteins (RIPs) are toxins with enzymatic activity rRNA- N-glycosidase. These proteins inhibit protein synthesis by specifically cleaving the adenine residue 4324 (A 4324 ) from ribosomal RNA 28S of the ribosomal subunit 60S. This process stops the protein synthesis, leading to...
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| Tipo de recurso: | tesis de maestría |
| Estado: | Versión publicada |
| Fecha de publicación: | 2019 |
| País: | Brasil |
| Institución: | Universidade Federal do Ceará (UFC) |
| Repositorio: | Repositório Institucional da Universidade Federal do Ceará (UFC) |
| Idioma: | portugués |
| OAI Identifier: | oai:repositorio.ufc.br:riufc/80224 |
| Acceso en línea: | http://repositorio.ufc.br/handle/riufc/80224 |
| Access Level: | acceso abierto |
| Palabra clave: | CNPQ::CIENCIAS BIOLOGICAS Proteínas inativadoras de ribossomos Jatropha podagrica Hook Podagrina Ribosome inactivation protein Podagrin |
| Sumario: | Ribosome inactivating proteins (RIPs) are toxins with enzymatic activity rRNA- N-glycosidase. These proteins inhibit protein synthesis by specifically cleaving the adenine residue 4324 (A 4324 ) from ribosomal RNA 28S of the ribosomal subunit 60S. This process stops the protein synthesis, leading to cell death. RIPs can be classified in type 1, 2 e 3. The main goal of this work was to isolate e characterize Jatropha podagrica Hook RIP and test its different in vitro biological activities. After collecting, cleaning the seeds, producing a flour and removing the lipids, the best protein extraction condition and purification protocols were determined. For the crude extracts (CE) the best buffer were Tris-HCl 20 mM pH 7,4. After that, a proteins precipitation with (NH 4 ) 2 SO 4 at 60% saturation was performed and denominated Fr0/60. The Fr0/60 was used for size exclusion chromatography (Sephadex G-100). After chromatography, it was obtained only one type 1 RIP, named podagrin. The podagrin showed rRNA-N-glycosidase activity and when submitted to dichroism circular analysis, podagrin presented a conformational pattern with predominance of sheet secondary structures. It was also performed with podagrin an anti-helminth assay, against Ascaridia galli. The most efficient concentration was 5 mg/mL of podagrin, with a 100% mortality of the parasite after 72 h. With these data, it is possible to conclude that podagrin is a type 1 RIP. |
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