A putative role for intramolecular regulatory mechanisms in the adaptor function of amphiphysin in endocytosis

Khashayar Farsad, Vladimir Slepnev, Giancarlo Ochoa, Laurie Daniell, Volker Hauke, Pietro De Camilli

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Amphiphysin 1 is a brain-specific protein enriched at the synapse and a major binding partner of several components of the clathrin-mediated endocytic machinery (Proc Natl Acad Sci USA 93 (1996) 331). It interacts with clathrin-coat proteins, dynamin, and membranes (Nat Cell Biol 1 (1999) 33; JBC). A role of amphiphysin in synaptic vesicle recycling is supported by both acute and chronic perturbation studies (Science 276 (1997) 259; Neuron 33 (2002) 789). Here we show that amphiphysin directly stimulates clathrin recruitment onto liposomes in an in vitro assay. Amphiphysin-dependent clathrin-coat recruitment is enhanced by the interaction of amphiphysin with dynamin. We also show that the amphiphysin SH3 domain binds full-length amphiphysin, likely via an internal poly-proline region, and that clathrin recruitment onto liposomes by amphiphysin is enhanced in the presence of the isolated amphiphysin SH3 domain. Expression of a mutant amphiphysin harboring two amino acid substitutions in the SH3 domain, and therefore unable to bind proline-containing motifs, induces an accumulation of large intracellular aggregates including amphiphysin, clathrin, AP-2, and other endocytic proteins, as well as a concomitant block of transferrin endocytosis. Thus, putative intramolecular interactions between the amphiphysin COOH-terminal SH3 domain and its internal poly-proline region may regulate clathrin recruitment onto membranes.

Original languageEnglish (US)
Pages (from-to)787-796
Number of pages10
JournalNeuropharmacology
Volume45
Issue number6
DOIs
StatePublished - Nov 2003
Externally publishedYes

Fingerprint

Endocytosis
Clathrin
src Homology Domains
Proline
Dynamins
Liposomes
amphiphysin
Synaptic Vesicles
Capsid Proteins
Amino Acid Substitution
Transferrin
Synapses
Proteins
Cell Membrane
Neurons

Keywords

  • Amphiphysin
  • Clathrin
  • Dynamin
  • Endocytosis

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Drug Discovery
  • Pharmacology

Cite this

A putative role for intramolecular regulatory mechanisms in the adaptor function of amphiphysin in endocytosis. / Farsad, Khashayar; Slepnev, Vladimir; Ochoa, Giancarlo; Daniell, Laurie; Hauke, Volker; De Camilli, Pietro.

In: Neuropharmacology, Vol. 45, No. 6, 11.2003, p. 787-796.

Research output: Contribution to journalArticle

Farsad, Khashayar ; Slepnev, Vladimir ; Ochoa, Giancarlo ; Daniell, Laurie ; Hauke, Volker ; De Camilli, Pietro. / A putative role for intramolecular regulatory mechanisms in the adaptor function of amphiphysin in endocytosis. In: Neuropharmacology. 2003 ; Vol. 45, No. 6. pp. 787-796.
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