Determining the conformational changes of multi-domain adaptor proteins driving clathrin mediated endocytosis

Clathrin-mediated endocytosis (CME) is driven by the assembly of adaptor proteins that participate in cargo selection, clathrin recruitment and membrane bending. Eps15, Dab2 and FCHo1/2 are early initiators of CME and they work together to recruit the adaptor proteins that drive vesicle formation. Multivalent interactions through both folded domains and intrinsically disordered regions of these proteins facilitate an interconnected protein network, that triggers the formation of biological condensates. However, the molecular details of
both the interactions and domain reorganization are still unkown. Using NMR spectroscopy, we have unveiled the local structure and the interaction modes of these proteins, suggesting the presence of long-range contacts affecting the global conformations. Therefore, we aim to understand the global conformational changes occurring within Eps15, FCHo1/2 and Dab2 proteins during the initiation of CME and how they relate to liquid-liquid phase separation.

网格蛋白介导的内吞作用（Clathrin-mediated endocytosis, CME）由参与货物选择、网格蛋白招募及膜弯曲的衔接蛋白组装所驱动。Eps15、Dab2与FCHo1/2是CME的早期起始因子，它们协同招募驱动囊泡形成的衔接蛋白。这些蛋白质通过折叠结构域与内在无序区域之间的多价相互作用，形成相互连接的蛋白质网络，进而触发生物凝聚体的形成。然而，相关相互作用与结构域重排的分子细节仍未明确。本研究借助核磁共振波谱法（NMR spectroscopy），揭示了这些蛋白质的局部结构与相互作用模式，表明存在影响其整体构象的长程相互作用。因此，本研究旨在阐明CME起始过程中Eps15、FCHo1/2及Dab2蛋白质的整体构象变化，及其与液液相分离的关联。