Epsin but not AP-2 supports reconstitution of endocytic clathrin-coated vesicles

• Verfasst von: Brod, Jan [VerfasserIn]
• Verfasst von: Hellwig, Andrea [VerfasserIn]
• Verfasst von: Wieland, Felix T. [VerfasserIn]
• Titel: Epsin but not AP-2 supports reconstitution of endocytic clathrin-coated vesicles
• Verf.angabe: Jan Brod, Andrea Hellwig and Felix T. Wieland
• Jahr: 2020
• Umfang: 13 S.
• Fussnoten: First published: 26 April 2020 ; Gesehen am 12.11.2021
• Titel Quelle: Enthalten in: Vereinigung der Europäischen Biochemischen GesellschaftenFEBS letters
• Ort Quelle: Chichester : Wiley, 1968
• Jahr Quelle: 2020
• Band/Heft Quelle: 594(2020), 14, Seite 2227-2239
• ISSN Quelle: 1873-3468
• DOI: doi:10.1002/1873-3468.13801
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: clathrin-coated vesicle
• Sach-SW: endocytosis
• Sach-SW: membrane scission
• Sach-SW: vesicle reconstitution
• Sach-SW: vesicular transport
• K10plus-PPN: 1777369487
• K10plus-PPN:
  Universitätsbibliothek
• Lokale URL UB: Zum Volltext

Abstract

Formation of clathrin-coated vesicles (CCVs) in receptor-mediated endocytosis is a mechanistically well-established process, in which clathrin, the adaptor protein complex AP-2, and the large GTPase dynamin play crucial roles. In order to obtain more mechanistic insight into this process, here we established a giant unilamellar vesicle (GUV)-based in vitro CCV reconstitution system with chemically defined components and the full-length recombinant proteins clathrin, AP-2, epsin-1, and dynamin-2. Our results support the predominant model in which hydrolysis of GTP by dynamin is a prerequisite to generate CCVs. Strikingly, in this system at near physiological concentrations of reagents, epsin-1 alone does not have the propensity for scission but is required for bud formation, whereas AP-2 and clathrin are not sufficient. Thus, our study reveals that epsin-1 is an important factor for the maturation of clathrin coated buds, a prerequisite for vesicle generation.