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Expression in Pichia pastoris of Ts5, a venom neurotoxin of Tityus serrulatus, with action on sodium channels

Grant number: 18/09876-0
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): September 01, 2018
Effective date (End): August 31, 2019
Field of knowledge:Health Sciences - Pharmacy
Principal Investigator:Eliane Candiani Arantes Braga
Grantee:Laura Martins Miranda Fernandes
Home Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

In Brazil, the species Tityus serrulatus scorpion has been widely studied, since it is responsible for severe poisonings in the country, with a high lethality rate. The venom of this species presents a very high toxicity and is the source of a diversity of molecules with potential biotechnological and pharmaceutical application, neurotoxins being the most important compounds due to their ability to interfere in the mechanisms of ionic permeability in excitable membrane cells and to interact specifically with voltage-dependent sodium and potassium channels. Thus, the study of the neurotoxins present in this venom has presented great potential for the pharmaceutical industry, seeking to develop new drugs, such as the case of Captopril. The objectives of this work are to standardize the heterologous expression of the Ts5 toxin in Pichia pastoris and perform the evaluation of the integrity of the recombinant toxin using heterologous protein expression techniques, purification from fast protein liquid chromatography (FPLC), gel electrophoresis and mass spectrometry by MALDI-TOF. Subsequently the results will be analyzed in order to assess whether the heterologous expression of the protein was effective, and whether the recombinant Ts5 maintains the integrity of the physiological activity of the native Ts5. This study represents unique importance in advancing the production of more efficient antivenoms and also in the study of new sources of medicines. In this way, this project shows to be of great relevance to the academic community, aiming to clarify the action of this toxin in the human immune system and the possible production of new drugs and antivenoms.