Centre for Host-Microbiome Interactions, Faculty of Dentistry, Oral & Craniofacial Sciences, King's College London, London, UK.
Department of Chemistry, King's College London, London, UK.
Methods Mol Biol. 2022;2542:163-176. 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.
2016 年，首次在人类真菌病原体中鉴定出一种肽毒素。它在白色念珠菌中被发现，并被命名为念珠菌溶素。念珠菌溶素是一种两亲性阳离子肽，可破坏细胞膜。与大多数溶细胞肽一样，念珠菌溶素有α-螺旋二级结构。由于念珠菌溶素的螺旋性和膜溶活性是致病性的关键因素，因此我们在这里描述了体外监测其膜溶功能和二级结构的方法。首先，通过直接电记录实时测量念珠菌溶素的膜通透性活性。其次，通过圆二色性光谱法测定念珠菌溶素的二级结构和螺旋性。这些生物物理方法为体外研究念珠菌溶素的活性和物理特性提供了一种手段，并将有助于确定念珠菌溶素和其他类似阳离子膜活性肽的结构和功能特征。
