Functional analysis of interaction sites on the N-terminal domain of clathrin heavy chain

Traffic. 2012 Jan;13(1):70-81. doi: 10.1111/j.1600-0854.2011.01289.x. Epub 2011 Oct 20.


In clathrin-mediated membrane traffic, clathrin does not bind directly to cargo and instead binds to adaptors that mediate this function. For endocytosis, the main adaptor is the adaptor protein (AP)-2 complex, but it is uncertain how clathrin contacts AP-2. Here we tested in human cells the importance of the three binding sites that have been identified so far on the N-terminal domain (NTD) of clathrin. We find that mutation of each of the three sites on the NTD, alone or in combination, does not block clathrin/AP-2-mediated endocytosis in the same way as deletion of the NTD. We report here the fourth and final site on the NTD that is required for clathrin/AP-2-mediated endocytic function. Each of the four interaction sites can operate alone to mediate endocytosis. The observed functional redundancy between interaction sites on the NTD explains how productivity of clathrin-coated vesicle formation is ensured.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Protein Complex 2 / genetics
  • Adaptor Protein Complex 2 / metabolism*
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Cell Membrane / metabolism*
  • Clathrin Heavy Chains / genetics
  • Clathrin Heavy Chains / metabolism*
  • Endocytosis*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Microscopy, Confocal
  • Models, Molecular
  • Molecular Sequence Data
  • Mutation
  • Protein Transport
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Sequence Alignment
  • Transfection


  • Adaptor Protein Complex 2
  • Recombinant Fusion Proteins
  • Clathrin Heavy Chains
  • Green Fluorescent Proteins