Structural characterization of proteins with action on ion channels by mass spectrometry

• Grant number: 12/16148-4
• Support Opportunities: Scholarships abroad - Research
• Start date: September 24, 2012
• End date: December 01, 2012
• Field of knowledge: Health Sciences - Pharmacy - Toxicological Analysis
• Principal Investigator: Eliane Candiani Arantes Braga
• Grantee: Eliane Candiani Arantes Braga
• Host Investigator: Edwin de Pauw
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: Université de Liège (ULg), Belgium

Abstract

Numerous studies performed with animal venoms have elucidated both the structure and the biological role of many venom components, and have shown the complex nature of venoms in both respects. The major constituents of scorpion venom are peptides and proteins with action on ion channels (Na+ or K+ channels) and the advent of matrix-assisted laser desorption-ionization (MALDI) and electrospray (ESI) as soft ionization technologies has permitted great strides in the study of proteinaceous toxins from scorpion venoms. Nowadays, mass spectrometry is the method of choice for the sequencing of peptides and proteins using soft ionization sources. After ionization, peptides are commonly sequenced by tandem mass spectrometry. The disulfide bonds are traditionally chemically reduced and alkylated before enzymatic digestion of the samples. This project proposes an internship at the Laboratory of Prof. Edwin De Pauw (Mass Spectrometry Laboratory, University Liege, Belgium) where mass spectrometry techniques are used for structural characterization of proteins and peptides. The technology will be transferred to the Laboratory Animal Toxins, which I am responsible, and therefore to FCFRP-USP, which already has a Central of Mass Spectrometry. The aim of this project is to learn to use mass spectrometry for structural characterization of proteins and peptides, with special attention to those that act on ion channels, which are targets of several studies of the Laboratory of Animal Toxins. To achieve these objectives, mass spectrometry assays will be performed to determine the molecular weight of peptides and proteins from scorpion venom, the de novo sequence of peptides, disulfide bonds and the presence of amidation at the C-terminal region. (AU)