2022 - In Vitro Biophysical Characterization of Candidalysin: A Fungal Peptide Toxin
Orbit 16 publication in Frontiers in Methods in Molecular Biology (2022), vol 2542
Lee S., Kichik N., Hepworth O., Richardson J., Naglik J.
Methods in Molecular Biology (2022) doi: 10.1007/978-1-0716-2549-1_12
In 2016, the first peptide toxin in any human fungal pathogen was identified. It was discovered in Candida albicans and was named candidalysin. Candidalysin is an amphipathic cationic peptide that damages cell membranes. Like most lytic peptides, candidalysin shows alpha-helical secondary structure. As the helicity and the membrane lytic activity of candidalysin are key factors for pathogenicity, here we describe in vitro approaches to monitor both its membrane-lytic function and the secondary structure. First, membrane permeabilization activity of candidalysin is measured in real time by direct electrical recording. Second, the secondary structure and helicity of candidalysin are determined by circular dichroism spectroscopy. These biophysical methods provide a means to characterize the activity and physical properties of candidalysin in vitro and will be useful in determining the structural and functional features of candidalysin and other similar cationic membrane-active peptides.