Novel binding sites on clathrin and adaptors regulate distinct aspects of coat assembly

Traffic. 2006 Dec;7(12):1688-700. doi: 10.1111/j.1600-0854.2006.00499.x. Epub 2006 Oct 17.

Abstract

Clathrin-coated vesicles (CCVs) sort proteins at the plasma membrane, endosomes and trans Golgi network for multiple membrane traffic pathways. Clathrin recruitment to membranes and its self-assembly into a polyhedral coat depends on adaptor molecules, which interact with membrane-associated vesicle cargo. To determine how adaptors induce clathrin recruitment and assembly, we mapped novel interaction sites between these coat components. A site in the ankle domain of the clathrin triskelion leg was identified that binds a common site on the appendages of tetrameric [AP1 and AP2] and monomeric (GGA1) adaptors. Mutagenesis and modeling studies suggested that the clathrin-GGA1 appendage interface is nonlinear, unlike other peptide-appendage interactions, but overlaps with a sandwich domain binding site for accessory protein peptides, allowing for competitive regulation of coated vesicle formation. A novel clathrin box in the GGA1 hinge region was also identified and shown to mediate membrane recruitment of clathrin, while disruption of the clathrin-GGA1 appendage interaction did not affect recruitment. Thus, the distinct sites for clathrin-adaptor interactions perform distinct functions, revealing new aspects to regulation of CCV formation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adaptor Proteins, Vesicular Transport / chemistry
  • Adaptor Proteins, Vesicular Transport / genetics
  • Adaptor Proteins, Vesicular Transport / metabolism*
  • Animals
  • Binding Sites
  • Clathrin / chemistry
  • Clathrin / metabolism*
  • Mice
  • Models, Molecular
  • Mutation / genetics
  • NIH 3T3 Cells
  • Protein Binding
  • Protein Structure, Quaternary
  • Protein Structure, Tertiary

Substances

  • Adaptor Proteins, Vesicular Transport
  • Clathrin