Clathrin-mediated endocytosis (CME) occurs via the formation of clathrin-coated vesicles from clathrin-coated pits (CCPs). Clathrin is recruited to CCPs through interactions between the AP2 complex and its N-terminal domain, which in turn recruits endocytic accessory proteins. Inhibitors of CME that interfere with clathrin function have been described, but their specificity and mechanisms of action are unclear. Here we show that overexpression of the N-terminal domain with (TDD) or without (TD) the distal leg inhibits CME and CCP dynamics by perturbing clathrin interactions with AP2 and SNX9. TDD overexpression does not affect clathrin-independent endocytosis or, surprisingly, AP1-dependent lysosomal trafficking from the Golgi. We designed small membrane–permeant peptides that encode key functional residues within the four known binding sites on the TD. One peptide, Wbox2, encoding residues along the W-box motif binding surface, binds to SNX9 and AP2 and potently and acutely inhibits CME.

中文翻译：

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Wbox2：网格蛋白末端结构域衍生的肽抑制剂，可抑制网格蛋白介导的内吞作用

网格蛋白介导的内吞作用 (CME) 通过从网格蛋白包被的凹坑 (CCP) 形成网格蛋白包被的囊泡而发生。网格蛋白通过 AP2 复合物与其 N 末端结构域之间的相互作用被招募到 CCP，进而招募内吞辅助蛋白。干扰网格蛋白功能的 CME 抑制剂已被描述，但其特异性和作用机制尚不清楚。在这里，我们表明，具有（TDD）或不具有（TD）远端腿的N端结构域的过度表达通过干扰网格蛋白与AP2和SNX9的相互作用来抑制CME和CCP动力学。TDD 过表达不影响网格蛋白独立的内吞作用，或者令人惊讶的是，不影响来自高尔基体的 AP1 依赖性溶酶体运输。我们设计了小的透膜肽，编码 TD 上四个已知结合位点内的关键功能残基。一种肽 Wbox2 编码沿 W-box 基序结合表面的残基，可与 SNX9 和 AP2 结合，并有效且急性地抑制 CME。