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Isolation and characterization of components with proteolytic activity presents in the fraction II-III from Tityus serrulatus venom

Grant number: 18/13110-2
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): October 01, 2018
Effective date (End): September 30, 2019
Field of knowledge:Health Sciences - Pharmacy
Principal Investigator:Eliane Candiani Arantes Braga
Grantee:Letícia Acosta
Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Scorpionism has a great relevance in Brazil, being responsible for most of the accidents involving venomous animals in the country and for causing several serious symptoms, which can lead to death. The most dangerous scorpion in the country is Tityus serrulatus, which venom has a wide range of components, including proteases (such as metalloproteases and serine proteases). To date, there are few studies on scorpion proteases; however, they may present important biotechnological and therapeutic applications. In addition, the study of these enzymes may contribute to increase knowledge about the venom composition and to the development of more efficient therapies for envenoming. Therefore, this project aims to isolate and characterize the components with proteolytic activity present in the fraction II-III from Tityus serrulatus venom. For this, first of all, several chromatographies will be performed to isolate the components present in the fraction of interest. Then, each of the peaks eluted from the chromatography will be subjected to an evaluation of the azocaseinolytic and fibrinogenolytic activities in the absence or presence of protease inhibitors (EDTA and PMSF) to identify the fractions that have serine proteases and metalloproteases and in which of them the proteolityc activity is more intense. The purity degree of each of these fractions will be evaluated by polyacrylamide gel electrophoresis with denaturing agent (SDS-PAGE) and the molecular mass of the components of these fractions will be determined by mass spectrometry (MALDI-TOF). This work may contribute to clarify the relevance of proteases to venom toxicity and to the post-translational modifications of their neurotoxins.