Chemistry Research Laboratory, Department of Chemistry, University of Oxford, UK.
Department of Chemistry, King's College London, UK.
FEBS J. 2023 Jan;290(2):428-441. doi: 10.1111/febs.16596. Epub 2022 Sep 19.
We exploit single-molecule tracking and optical single channel recording in droplet interface bilayers to resolve the assembly pathway and pore formation of the archetypical cholesterol-dependent cytolysin nanopore, Perfringolysin O. We follow the stoichiometry and diffusion of Perfringolysin O complexes during assembly with 60 ms temporal resolution and 20 nm spatial precision. Our results suggest individual nascent complexes can insert into the lipid membrane where they continue active assembly. Overall, these data support a model of stepwise irreversible assembly dominated by monomer addition, but with infrequent assembly from larger partial complexes.
我们利用单分子跟踪和液滴界面双层中的光学单通道记录来解析原型胆固醇依赖性细胞溶素纳米孔，即产气荚膜梭菌溶菌素 O 的组装途径和孔形成。我们以 60ms 的时间分辨率和 20nm 的空间精度来跟踪产气荚膜梭菌溶菌素 O 复合物在组装过程中的化学计量和扩散。我们的结果表明，单个新生复合物可以插入脂质膜中，在那里它们继续进行活跃的组装。总的来说，这些数据支持一个逐步不可逆组装的模型，该模型主要由单体添加主导，但也有来自较大部分复合物的偶发组装。
