2. Academic Validation
  3. Structure of the endocytic adaptor complex reveals the basis for efficient membrane anchoring during clathrin-mediated endocytosis

Structure of the endocytic adaptor complex reveals the basis for efficient membrane anchoring during clathrin-mediated endocytosis

  • Nat Commun. 2021 May 17;12(1):2889. doi: 10.1038/s41467-021-23151-7.
Javier Lizarrondo # 1,  David P Klebl # 2,  Stephan Niebling 1,  Marc Abella 3,  Martin A Schroer 1,  Haydyn D T Mertens 1,  Katharina Veith 1,  Roland Thuenauer 4,  Dmitri I Svergun 1,  Michal Skruzny 3,  Frank Sobott 5 6,  Stephen P Muench 2,  Maria M Garcia-Alai 7 8
Affiliations

Affiliations

  • 1 European Molecular Biology Laboratory, Hamburg Outstation, Hamburg, Germany.
  • 2 School of Biomedical Sciences, Faculty of Biological Sciences and Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, UK.
  • 3 Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology and LOEWE Center for Synthetic Microbiology (SYNMIKRO), Marburg, Germany.
  • 4 Technology Platform Microscopy and Image Analysis, Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
  • 5 School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, UK.
  • 6 Department of Chemistry, Biomolecular and Analytical Mass Spectrometry group, University of Antwerp, Antwerp, Belgium.
  • 7 European Molecular Biology Laboratory, Hamburg Outstation, Hamburg, Germany. garcia@embl-hamburg.de.
  • 8 Centre for Structural Systems Biology, Hamburg, Germany. garcia@embl-hamburg.de.
  • # Contributed equally.
PMID: 34001871 DOI: 10.1038/s41467-021-23151-7
Abstract

During clathrin-mediated endocytosis, a complex and dynamic network of protein-membrane interactions cooperate to achieve membrane invagination. Throughout this process in yeast, endocytic coat adaptors, Sla2 and Ent1, must remain attached to the plasma membrane to transmit force from the actin Cytoskeleton required for successful membrane invagination. Here, we present a cryo-EM structure of a 16-mer complex of the ANTH and ENTH membrane-binding domains from Sla2 and Ent1 bound to PIP2 that constitutes the anchor to the plasma membrane. Detailed in vitro and in vivo mutagenesis of the complex interfaces delineate the key interactions for complex formation and deficient cell growth phenotypes demonstrate its biological relevance. A hetero-tetrameric unit binds PIP2 molecules at the ANTH-ENTH interfaces and can form larger assemblies to contribute to membrane remodeling. Finally, a time-resolved small-angle X-ray scattering study of the interaction of these adaptor domains in vitro suggests that ANTH and ENTH domains have evolved to achieve a fast subsecond timescale assembly in the presence of PIP2 and do not require further proteins to form a stable complex. Together, these findings provide a molecular understanding of an essential piece in the molecular puzzle of clathrin-coated endocytic sites.

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