| Grant number: | 14/06170-8 |
| Support Opportunities: | Scholarships in Brazil - Master |
| Start date: | October 01, 2014 |
| End date: | July 31, 2016 |
| Field of knowledge: | Health Sciences - Pharmacy - Toxicological Analysis |
| Agreement: | Coordination of Improvement of Higher Education Personnel (CAPES) |
| Principal Investigator: | Eliane Candiani Arantes Braga |
| Grantee: | Gisele Adriano Wiezel |
| Host Institution: | Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil |
| Associated research grant: | 11/23236-4 - Native and recombinant animal toxins: functional, structural and molecular analysis, AP.TEM |
| Associated scholarship(s): | 14/23285-3 - Characterization of post-translation modifications of an L-amino acid oxidase isolated from Crotalus durissus terrificus snake venom, BE.EP.MS |
Abstract Snakebites represent a serious public health problem in tropical countries. In Brazil, the vast majority of reported accidents are caused by Viperidae family and genus Crotalus is responsible for the higher mortality of the accidents. Snake venoms are complex mixtures of proteins, peptides and organic compounds that affect a huge variety of biological process. L-amino acid oxidase (LAAO) is an enzyme present in these venoms which catalyses L-amino acids oxidation and produce ±-ketoacids, ammonia and hydrogen peroxide. Various functional activities of LAAOs have been reported and it has been demonstrated and attractive to biotechnology area. Among them, antitumor and antimicrobial activities, activity on blood coagulation, anti-HIV activity, edema induction and also modulation of system immune cells can be cited. Many authors have reported action of L-amino acid oxidases against parasites from Leishmania genus, including L. braziliensis, L. amazonensis, L. donovani and others. Leishmaniasis is considered a public health problem and, nowadays, it has been gaining great interest mainly due to the expansion of international travels and population growth. In South America, Leishmaniasis is an endemic tropical disease and it has few therapeutic approaches. Therefore, considering LAAO action against Leishmania spp, the importance of Leishmaniasis to public health and the biotechnological potential of these enzymes, it is intended to isolate and characterize pioneering an LAAO present in C. d. terrificus venom, aiming to increase the knowledge about the mechanism of action of this enzyme, its relevance in the immune response observed in poisoning and especially its possible application as a therapeutic agent and/or adjuvant for the treatment of Leishmaniasis. Isolation of LAAO will be accomplished through fast protein liquid chromatography and its characterization will be taken from the sequencing of its internal peptides, determination of molar mass spectrometry, evaluation of specificity against different substrates, determination of kinetic parameters (Km, Vmax, kcat) and in vitro evaluation of immune response induced in fibroblasts by analyzing the cytotoxicity and dosage of cytokines, nitric oxide and lipid mediator. Leishmanicidal activity will also be assessed. (AU) | |
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