STRUCTURAL AND MECHANISTIC INSIGHTS INTO PORE FORMATION BY PROTEINS OF THE MEMBRANE ATTACK COMPLEX/PERFORIN SUPERFAMILY

孔形成蛋白在介导宿主病原体相互作用中起重要作用。它们存在于细菌和真核生物中，涵盖免疫效应物、毒素和致病毒力因子。成孔蛋白的膜攻击复合物/穿孔素（MACPF）超家族的成员的特征在于能够刺穿脂质膜的共同三维折叠。该项目旨在推导出含MACPF蛋白如何形成孔的分子机制，以及这一过程如何受到调节蛋白和局部脂质环境的影响。它将探索免疫效应器，膜攻击复合物（MAC）的分子动力学模拟，结合其抑制剂CD 59，以确定蛋白质-脂质相互作用，并探索膜厚度的变化，这可能是CD 59的招募信号。在这些数据的基础上，该项目还将研究细菌毒力因子如何协同CD 59形成核孔。胆固醇依赖性溶细胞素（CDC）是细菌成孔蛋白的广泛家族，其中一个子集使用人细胞上的CD 59作为其孔形成途径的一部分。在这里，我们将使用生物化学，结构和计算方法的组合来了解它们的成孔机制。

Pore forming proteins play an important role in mediating host pathogen interactions. They are found in bacteria and eukaryotes, and span immune effectors, toxins, and pathogenic virulence factors. Members of the membrane attack complex/perforin (MACPF) superfamily of pore-forming proteins are characterised by a common three-dimensional fold able to puncture lipid membranes. This project aims to derive a molecular mechanism for how MACPF-containing proteins form pores and how this process is influenced by regulator proteins and the local lipid environment. It will explore molecular dynamics simulations of the immune effector, membrane attack complex (MAC), bound to its inhibitor CD59 to identify protein-lipid interactions and explore changes in membrane thickness, which may serve as CD59 recruitment signals. Building on these data, this project will also investigate how bacterial virulence factors co-opt CD59 to nucleate pore formation. Cholesterol-dependent cytolysins (CDCs) are a broad family of bacterial pore-forming proteins, a subset of which use CD59 on human cells as part of their pore formation pathway. Here we will use a combination of biochemical, structural, and computational approaches to understand their pore-forming mechanism.