Characterization of Ts20, a new protein likely to be of biotechnological interest in the Tityus serrulatus venom

• Grant number: 20/09434-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: November 01, 2020
• End date: October 31, 2021
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Eliane Candiani Arantes Braga
• Grantee: Marco Cezar Camargo Carmona
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Scorpionism has great relevance in Brazil, being responsible for the majority of accidents involving venomous animals in the country and for causing several serious symptoms, which can lead to death. The scorpion species Tityus serrulatus represents a serious threat to human health in Brazil, it is among the animals that produce the most dangerous venoms for mammals representing the highest percentage of accidents. The venom composition has a wide range of components, mainly neurotoxins with action on ion channels and several compounds that have not yet been isolated and / or studied. Therefore, this work aims to purify and characterize a new protein present in fraction 45 obtained from the reverse phase chromatography (C-18) of the Tityus serrulatus venom, called Ts20. For this, first, the venom will be fractionated by fast protein liquid chromatography (Fast Protein Liquid Chromatography - FPLC), in a C-18 reverse phase column. Then, the fraction of interest (Fraction 45) will be subjected to a polyacrylamide gel electrophoresis to verify its degree of purity. Subsequently, amino terminal protein sequencing will be performed by Edman degradation and its molecular mass will be determined by mass spectrometry (MALDI-TOF). To characterize it functionally, Ts20 will be subjected to tests to detect possible cytotoxic activities, using the MTT technique, and antiviral in vitro. Additionally, its ability to inhibit proteases will be evaluated. This work may contribute to clarify the relevance of this protein in venom, contributing to a better understanding of the picture of poisoning by the scorpion Tityus serrulatus, as well as allowing to evaluate the potential of Ts20 as a new molecule with antiviral activity.