Clathrin-mediated endocytosis is an essential cellular internalization pathway involving the dynamic assembly of clathrin and accessory proteins to form membrane-bound vesicles. The evolutionarily ancient TSET–TPLATE complex (TPC) plays an essential, but ill-defined role in endocytosis in plants. Here we show that two highly disordered TPC subunits, AtEH1 and AtEH2, function as scaffolds to drive biomolecular condensation of the complex. These condensates specifically nucleate on the plasma membrane through interactions with anionic phospholipids, and facilitate the dynamic recruitment and assembly of clathrin, as well as early- and late-stage endocytic accessory proteins. Importantly, condensation promotes ordered clathrin assemblies. TPC-driven biomolecular condensation thereby facilitates dynamic protein assemblies throughout clathrin-mediated endocytosis. Furthermore, we show that a disordered region of AtEH1 controls the material properties of endocytic condensates in vivo. Alteration of these material properties disturbs the recruitment of accessory proteins, influences endocytosis dynamics and impairs plant responsiveness. Our findings reveal how collective interactions shape endocytosis.

网格蛋白介导的内吞作用是一种重要的细胞内化途径，涉及网格蛋白和辅助蛋白的动态组装以形成膜结合囊泡。进化上古老的 TSET-TPLATE 复合物（TPC）在植物的内吞作用中发挥着重要但不明确的作用。在这里，我们展示了两个高度无序的 TPC 亚基 AtEH1 和 AtEH2，作为支架驱动复合物的生物分子缩合。这些缩合物通过与阴离子磷脂的相互作用在质膜上特异性成核，并促进网格蛋白以及早期和晚期内吞辅助蛋白的动态募集和组装。重要的是，缩合促进了网格蛋白有序组装。 TPC 驱动的生物分子缩合从而促进了网格蛋白介导的内吞作用中的动态蛋白质组装。此外，我们还发现 AtEH1 的无序区域控制着体内内吞凝聚物的材料特性。这些材料特性的改变会干扰辅助蛋白的募集，影响内吞动力学并损害植物反应性。我们的研究结果揭示了集体相互作用如何塑造内吞作用。