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

孔道形成蛋白在介导宿主病原菌相互作用中起着重要作用。它们存在于细菌和真核生物中，跨越免疫效应物、毒素和致病毒力因子。膜攻击复合体/穿孔素(MACPF)超家族的成孔蛋白具有一个共同的三维折叠结构，能够穿透脂质膜。这个项目的目的是找出含有MACPF的蛋白质如何形成毛孔的分子机制，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-膜攻击复合体(MAC)与其抑制剂CD59结合的分子动力学模拟，以确定蛋白质与脂质的相互作用，并探索膜厚度的变化，这可能作为CD59募集信号。在这些数据的基础上，该项目还将调查细菌毒力因子如何协同CD59形成毛孔。胆固醇依赖细胞溶蛋白(CDCs)是细菌致孔蛋白的一个广泛家族，其中一部分利用人类细胞上的CD59作为其孔道的一部分。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孔形成机制。

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.