Clathrin interacts specifically with amphiphysin and is displaced by dynamin

• Published in: FEBS letters 1997-08, Vol.413 (2), p.319-322
• Main Authors: McMahon, Harvey T, Wigge, Patrick, Smith, Corrin
• Format: Article
• Language: English
• Subjects: Adaptor Protein Complex alpha Subunits; Adaptor Proteins, Vesicular Transport; Amphiphysin; Animals; Brain - metabolism; Cell Extracts; Clathrin; Clathrin - metabolism; Coated pit; Dynamin; Dynamin I; Dynamins; GTP Phosphohydrolases - metabolism; Membrane Proteins - metabolism; Molecular Sequence Data; Nerve Tissue Proteins - metabolism; Protein Binding; Rats; Recombinant Fusion Proteins; Swine; Synaptic vesicle Endocytosis
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(97)00928-9

Amphiphysin is an SH3 domain protein that has been implicated in synaptic vesicle endocytosis. We have recently cloned a second amphiphysin isoform, Amph2 (sequence submitted to GenBank, Y13380). Proteins capable of forming a complex with amphiphysin were isolated from rat brain by using recombinant GST-Amph2 for binding experiments. As well as interacting with dynamin I, the full-length protein bound to a weaker 180-kDa band. Immunoblotting demonstrated this protein to be clathrin. To address whether this is a direct interaction, the clathrin binding to amphiphysin was reconstituted in vitro with purified proteins. The N-terminal domain of Amph2 is sufficient for clathrin binding. Dynamin, which interacts with the SH3 domain of Amph2, displaces clathrin from the N-terminus. We propose a model that may explain how clathrin and dynamin are recruited to non-overlapping sites of the coated pit.