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Biochemical and in vitro evaluation of fibroblast activation and leishmanicidal potential of an L-amino acid oxidase (LAAO) from Crotalus durissus terrificus venom

Grant number: 14/06170-8
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): October 01, 2014
Effective date (End): July 31, 2016
Field of knowledge:Health Sciences - Pharmacy - Toxicological Analysis
Acordo de Cooperação: 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


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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Scientific publications (4)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
WIEZEL, GISELE A.; RUSTIGUEL, JOANE K.; MORGENSTERN, DAVID; ZOCCAL, KARINA E.; FACCIOLI, LUCIA H.; CRISTINA NONATO, M.; UEBERHEIDE, BEATRIX; ARANTES, ELIANE C.. Insights into the structure, function and stability of bordonein-L, the first L-amino acid oxidase from Crotalus durissus terrificus snake venom. Biochimie, v. 163, p. 33-49, . (14/03332-7, 11/23236-4, 15/24010-0, 14/23285-3, 14/06170-8, 14/07125-6)
BOLDRINI-FRANCA, JOHARA; COLOGNA, CAMILA TAKENO; PUCCA, MANUELA BERTO; FIGUEIREDO BORDON, KARLA DE CASTRO; AMORIM, FERNANDO GOBBI; PINO ANJOLETTE, FERNANDO ANTONIO; CORDEIRO, FRANCIELLE ALMEIDA; WIEZEL, GISELE ADRIANO; CERNI, FELIPE AUGUSTO; PINHEIRO-JUNIOR, ERNESTO LOPES; et al. Minor snake venom proteins: Structure, function and potential applications. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, v. 1861, n. 4, p. 824-838, . (14/06170-8, 11/12317-3, 11/23236-4, 15/16714-8, 12/13590-8, 13/26619-7, 14/15644-3, 12/12954-6, 13/26200-6)
BORDON, KARLA C. F.; WIEZEL, GISELE A.; AMORIM, FERNANDA G.; ARANTES, ELIANE C.. Arthropod venom Hyaluronidases: biochemical properties and potential applications in medicine and biotechnology. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 21, . (11/12317-3, 14/06170-8, 11/23236-4)
BORDON, KARLA C. F.; WIEZEL, GISELE A.; CABRAL, HAMILTON; ARANTES, ELIANE C.. Bordonein-L, a new L-amino acid oxidase from Crotalus durissus terrificus snake venom: isolation, preliminary characterization and enzyme stability. Journal of Venomous Animals and Toxins including Tropical Diseases, v. 21, . (14/06170-8, 11/23236-4)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
WIEZEL, Gisele Adriano. Biochemical characterization and in vitro evaluation of the fibroblast activation and the leishmanicide potential of an L-amino acid oxidase (LAAO) from Crotalus durissus terrificus venom. 2016. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC) Ribeirão Preto.

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