Sínteses e estudos dos diferentes peptídeos de fusão dos flavivirus causadores da dengue

• Ano de defesa: 2013
• Autor(a) principal: Cespedes, Graziely Ferreira [UNESP]
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual Paulista (Unesp)
• Palavras-chave em Português: Biotecnologia; Peptídeos; Fluorescencia; Dengue; Peptides
• Link de acesso: http://hdl.handle.net/11449/108473

Dengue virus affects millions of people worldwide every year. The process of viral infection via endosomal is mediated by the sequence 98-112 (DRGWGNGCGLFGKGG), known as the fusion peptide. This segment is part of the glycoprotein envelope of the virus. All flavivirus, including all four dengue serotypes, contain this fusion sequence and all serotypes present high sequence identity. Although intense researches, the dengue cases have increased and, up to date, there are no specific antiviral drugs to combat this disease. Thus, understanding the virus interaction with host cells is essential for the search and development of new drugs against the dengue virus. Therefore, this study craved a better understanding of the mode of action of the dengue fusion peptide sequence and the importance of its lateral regions. This project aimed to assess the contributions of the lateral regions of the fusion peptide (88-97 and 113-123 sequences) on its activity, for the four dengue serotypes. The synthesis of peptides was performed by solid phase peptide synthesis (SPPS). The membrane models used were zwitterionic micelles (LPC) and vesicles (egg PC and POPG). Since the exposure of this sequence and the resulting membrane fusion is dependent on pH, studies were performed in pH 5.5 and 7.1. In these studies, the techniques of Resonance Energy Transfer Föster (FRET), circular dichroism, fluorescence, Langmuir monolayers and BAM (Brewster angle microscopy) were used. The amino acid and natural fluorophore tryptophan was used as a probe to analyze the insertion depth of the fusion peptide in the membrane mimetic by fluorescence. The results showed that the SPPS was feasible, obtaining the desired materials. Assessing the fusogenic ability of 14 peptides studied, the data indicated that each serotype has different activity and the serotype 2 peptides presented the major interaction. The studies of interaction peptide/membrane mimetic indicated...