Next-generation small molecule inhibitors of clathrin function acutely inhibit endocytosis

IF 4.6 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Structure Pub Date : 2025-03-19 DOI:10.1016/j.str.2025.02.011

André Horatscheck, Michael Krauß, Haydar Bulut, Valerie Chambon, Massilullah Shafaq Zadah, Estelle Dransart, Kimberly Peloza, Karine F. Santos, Mark J. Robertson, Kate Prichard, Sandra Miksche, Silke Radetzki, Jens-Peter von Kries, Markus C. Wahl, Adam McCluskey, Ludger Johannes, Volker Haucke, Marc Nazaré

{"title":"Next-generation small molecule inhibitors of clathrin function acutely inhibit endocytosis","authors":"André Horatscheck, Michael Krauß, Haydar Bulut, Valerie Chambon, Massilullah Shafaq Zadah, Estelle Dransart, Kimberly Peloza, Karine F. Santos, Mark J. Robertson, Kate Prichard, Sandra Miksche, Silke Radetzki, Jens-Peter von Kries, Markus C. Wahl, Adam McCluskey, Ludger Johannes, Volker Haucke, Marc Nazaré","doi":"10.1016/j.str.2025.02.011","DOIUrl":null,"url":null,"abstract":"Clathrin-mediated endocytosis (CME) is the predominant endocytic pathway in eukaryotic cells and a major regulator of cell physiology as it facilitates the internalization of receptors, channels, and transporters and viral entry. The clathrin terminal domain acts as a central protein interaction hub within the endocytic protein network. Previously described inhibitors of CME display off-target activities that result in cytotoxicity, providing limitations to their use. We report the development and characterization of next-generation small molecule inhibitors of clathrin terminal domain function. These compounds termed Pitstop 2c and Pitstop 2d occupy the binding site within the clathrin terminal domain for endocytic protein ligands including epsin, resulting in potent inhibition of receptor-mediated endocytosis and reduced entry of vesicular stomatitis virus (VSV) with minimal cytotoxic side effects. Next-generation Pitstops thus provide an improved toolset to address clathrin function in cell physiology with potential applications as inhibitors of virus and pathogen entry.","PeriodicalId":22168,"journal":{"name":"Structure","volume":"32 1","pages":""},"PeriodicalIF":4.6000,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Structure","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1016/j.str.2025.02.011","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Clathrin-mediated endocytosis (CME) is the predominant endocytic pathway in eukaryotic cells and a major regulator of cell physiology as it facilitates the internalization of receptors, channels, and transporters and viral entry. The clathrin terminal domain acts as a central protein interaction hub within the endocytic protein network. Previously described inhibitors of CME display off-target activities that result in cytotoxicity, providing limitations to their use. We report the development and characterization of next-generation small molecule inhibitors of clathrin terminal domain function. These compounds termed Pitstop 2c and Pitstop 2d occupy the binding site within the clathrin terminal domain for endocytic protein ligands including epsin, resulting in potent inhibition of receptor-mediated endocytosis and reduced entry of vesicular stomatitis virus (VSV) with minimal cytotoxic side effects. Next-generation Pitstops thus provide an improved toolset to address clathrin function in cell physiology with potential applications as inhibitors of virus and pathogen entry.

Abstract Image

查看原文

微信好友朋友圈 QQ好友复制链接

本刊更多论文

新一代小分子网格蛋白功能抑制剂急性抑制内吞作用

网格蛋白介导的内吞作用（CME）是真核细胞中主要的内吞途径，也是细胞生理的主要调节因子，因为它促进了受体、通道和转运体的内化以及病毒的进入。网格蛋白末端结构域在内吞蛋白网络中起着中心蛋白相互作用枢纽的作用。先前描述的CME抑制剂显示脱靶活性，导致细胞毒性，限制了它们的使用。我们报道了下一代网格蛋白末端结构域功能小分子抑制剂的开发和表征。这些被称为Pitstop 2c和Pitstop 2d的化合物占据了包括epsin在内的内吞蛋白配体的网格蛋白末端区域的结合位点，从而有效抑制受体介导的内吞作用，减少水疱性口炎病毒（VSV）的进入，并且细胞毒性副作用最小。因此，下一代维修站提供了一种改进的工具集来解决网格蛋白在细胞生理学中的功能，并具有作为病毒和病原体进入抑制剂的潜在应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文去求助

来源期刊

Structure 生物-生化与分子生物学

CiteScore

8.90

自引率

1.80%

发文量

155

审稿时长

3-8 weeks

期刊介绍： Structure aims to publish papers of exceptional interest in the field of structural biology. The journal strives to be essential reading for structural biologists, as well as biologists and biochemists that are interested in macromolecular structure and function. Structure strongly encourages the submission of manuscripts that present structural and molecular insights into biological function and mechanism. Other reports that address fundamental questions in structural biology, such as structure-based examinations of protein evolution, folding, and/or design, will also be considered. We will consider the application of any method, experimental or computational, at high or low resolution, to conduct structural investigations, as long as the method is appropriate for the biological, functional, and mechanistic question(s) being addressed. Likewise, reports describing single-molecule analysis of biological mechanisms are welcome. In general, the editors encourage submission of experimental structural studies that are enriched by an analysis of structure-activity relationships and will not consider studies that solely report structural information unless the structure or analysis is of exceptional and broad interest. Studies reporting only homology models, de novo models, or molecular dynamics simulations are also discouraged unless the models are informed by or validated by novel experimental data; rationalization of a large body of existing experimental evidence and making testable predictions based on a model or simulation is often not considered sufficient.