Exploring Host Defense in Xenopus laevis by Harvesting proteins and peptides from Frog Tissue Using Trizol Extraction and Analysis via Tandem Mass Spectrometry

Abstract

This project investigates antimicrobial peptides (AMPs), cytokines, and chemokines present in Xenopus laevis (African clawed frog) skin tissue, using Trizol for protein extraction and Tandem Mass Spectrometry (MS/MS) results to identify AMPs. Antimicrobial peptides are small proteins consisting of short chains of amino acids that can have antibacterial, antiviral, antifungal, anticancer, and other biological activities. Studying immunity in frogs is important because immunity in amphibians is poorly understood and has not been investigated as thoroughly as in other animals. Most of the research conducted on frogs to date has focused on their skin secretions rather than frog tissue. This research will contribute to our fundamental understanding of immunity in frogs and may be applicable to humans’ own host defense systems, as well. In this study, Trizol and bead beating are compared to see which method of protein extraction works best for frog tissue. Once the method of protein extraction from tissue has been established, C8 and C18 columns will be compared during the solid phase extraction portion of the protocol. After analyzing the resulting samples using tandem mass spectrometry, the data will be processed to determine the proteins and peptides that are present in the tissue samples.