Biochemical, structural and functional characterization of an L-amino acid oxidase from the yellow venom of the rattlesnake Crotalus durissus terrificus

• Grant number: 11/12267-6
• Support Opportunities: Regular Research Grants
• Start date: January 01, 2012
• End date: June 30, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Maurício Ventura Mazzi
• Grantee: Maurício Ventura Mazzi
• Host Institution: Pró-Reitoria de Pós-Graduação e Pesquisa. Centro Universitário Hermínio Ometto (UNIARARAS). Fundação Hermínio Ometto (FHO). Araras , SP, Brazil

Abstract

Proteins and peptides found in the venom of poisonous animals have contributed significantly to the development of Biological and Biomedical Sciences, as well as Biotechnology. Considering marked structural similarities between many of these biomolecules and some peptides and proteins found in other organisms, which enables them to interact specifically within biological systems, such toxins may contribute to the understanding of physiological processes, or may serve as prototypes for therapeutic agents. While venom constituents are generally unsuitable for direct clinical use, a rational design approach and structural modification of specific chemical moieties, based on the elucidation of their native structure and biological activities, enables such toxins to be employed as therapeutic agents in many diseases. The crotalic rattlesnake venom is rich in enzymes and peptides, which trigger important effects on skeletal muscles, nervous system, kidneys and blood clotting system. Crotalic venom components include the most studied proteins crotoxin, crotamin, gyroxin, convulxin and thrombin-like enzyme. In addition, other less studied protein components, such as L-amino acid oxidases (LAAOs), have potent cytotoxic and antimicrobial effects, which make them important pharmacological models for the design of new antibiotics. LAAOs are flavoproteins that confer a yellow hue to the snake venom, and have been triggering a variety of adverse effects, such as platelet aggregation, hemorrhage, edema, cytotoxicity and apoptosis induction, as well as a wide spectrum of bactericidal and anti-parasitic activities. Besides LAAOs pharmacological properties and attractive biotechnological potential, it is known that these enzymes can differ significantly in structure and substrate preferences. Thus, the aim of this project is to isolate and study the structure, as well as the biochemical and pharmacological activities of a LAAO identified in the yellow venom of the snake Crotalus durissus terrificus. This enzyme will be isolated and purified from the venom by chromatographic techniques, and its biochemical properties will be evaluated by N-terminal sequencing, X-ray diffraction crystallography, and enzyme kinetics assay at different conditions and substrates. We also propose to evaluate some LAAO biological properties, such as hemorrhagic, myotoxic, coagulant, anticoagulant and antibiotic activities. Results obtained in this study will certainly contribute for revealing the structure-function relationship of such toxin, as well as their possible applications in basic and applied research. (AU)

Scientific publications

(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)

TEIXEIRA, TUILA LEVEGHIM; OLIVEIRA SILVA, VIVIANE ALINE; DA CUNHA, DANIEL BATISTA; POLETTINI, FLAVIA LINO; THOMAZ, CAMILA DANIELE; PIANCA, ARIANA APARECIDA; ZAMBOM, FABIANA LETICIA; DA SILVA LEITAO MAZZI, DENISE PIMENTA; REIS, RUI MANUEL; MAZZI, MAURICIO VENTURA. Isolation, characterization and screening of the in vitro cytotoxic activity of a novel L-amino acid oxidase (LAAOcdt) from Crotalus durissus terrificus venom on human cancer cell lines. Toxicon, v. 119, p. 203-217, SEP 1 2016. (11/12267-6)