Next-generation small molecule inhibitors of clathrin function acutely inhibit endocytosis
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.
Authors
- M.C. Wahl (ORCID: https://orcid.org/0000-0002-2811-5307)
- Sandra Miksche
- Volker Haucke (ORCID: https://orcid.org/0000-0003-3119-6993)
- Kate Prichard (ORCID: https://orcid.org/0000-0003-0458-5037)
- M. Krauß (ORCID: https://orcid.org/0000-0003-1010-9723)
- Kimberly Peloza (ORCID: https://orcid.org/0000-0002-8255-5311)
- Karine Santos (ORCID: https://orcid.org/0000-0003-0467-7708)
- Adam McCluskey (ORCID: https://orcid.org/0000-0001-7125-863X)
- Silke Radetzki (ORCID: https://orcid.org/0000-0003-2688-2868)
- Haydar Bulut (ORCID: https://orcid.org/0000-0003-0262-6296)
- Estelle Dransart (ORCID: https://orcid.org/0000-0003-2547-6825)
- Marc Nazaré (ORCID: https://orcid.org/0000-0002-1602-2330)
- Valérie Chambon
- Mark J. Robertson (ORCID: https://orcid.org/0000-0002-2140-2717)
- Jens Peter von Kries (ORCID: https://orcid.org/0000-0003-4716-4988)
- Ludger Johannes (ORCID: https://orcid.org/0000-0002-2168-0004)
- André Horatscheck (ORCID: https://orcid.org/0000-0001-6485-6391)
- Massilullah Shafaq Zadah
Institutions
- Centre National de la Recherche Scientifique (FR)
- Inserm (FR)
- Université Paris Sciences et Lettres (FR)
- Helmholtz-Zentrum Berlin für Materialien und Energie (DE)
- Leibniz-Forschungsinstitut für Molekulare Pharmakologie (DE)
- Freie Universität Berlin (DE)
- University of Newcastle Australia (AU)
- Institut Curie (FR)
Publication Details
- Journal
- Structure
- Published
- 2025-03-19
- DOI
- https://doi.org/10.1016/j.str.2025.02.011
- Citations
- 6
- Primary Topic
- Cellular transport and secretion
- Type
- article
- Field-Weighted Citation Impact
- 6.91