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A conserved clathrin assembly motif essential for synaptic vesicle endocytosis.
- Source :
-
The Journal of neuroscience : the official journal of the Society for Neuroscience [J Neurosci] 2000 Dec 01; Vol. 20 (23), pp. 8667-76. - Publication Year :
- 2000
-
Abstract
- Although clathrin assembly by adaptor proteins (APs) plays a major role in the recycling of synaptic vesicles, the molecular mechanism that allows APs to assemble clathrin is poorly understood. Here we demonstrate that AP180, like AP-2 and AP-3, binds to the N-terminal domain of clathrin. Sequence analysis reveals a motif, containing the sequence DLL, that exists in multiple copies in many clathrin APs. Progressive deletion of these motifs caused a gradual reduction in the ability of AP180 to assemble clathrin in vitro. Peptides from AP180 or AP-2 containing this motif also competitively inhibited clathrin assembly by either protein. Microinjection of these peptides into squid giant presynaptic terminals reversibly blocked synaptic transmission and inhibited synaptic vesicle endocytosis by preventing coated pit formation at the plasma membrane. These results indicate that the DLL motif confers clathrin assembly properties to AP180 and AP-2 and, perhaps, to other APs. We propose that APs promote clathrin assembly by cross-linking clathrin triskelia via multivalent interactions between repeated DLL motifs in the APs and complementary binding sites on the N-terminal domain of clathrin. These results reveal the structural basis for clathrin assembly and provide novel insights into the molecular mechanism of clathrin-mediated synaptic vesicle endocytosis.
- Subjects :
- Adaptor Protein Complex alpha Subunits
Adaptor Proteins, Vesicular Transport
Animals
Binding Sites genetics
Binding, Competitive drug effects
Binding, Competitive genetics
Cattle
Cells, Cultured
Clathrin chemistry
Clathrin genetics
Coated Pits, Cell-Membrane drug effects
Coated Pits, Cell-Membrane metabolism
Decapodiformes
Endocytosis drug effects
Membrane Proteins genetics
Membrane Proteins metabolism
Microinjections
Nerve Tissue Proteins genetics
Peptides administration & dosage
Peptides chemistry
Peptides genetics
Phosphoproteins genetics
Presynaptic Terminals drug effects
Presynaptic Terminals metabolism
Rats
Surface Plasmon Resonance
Synaptic Transmission drug effects
Synaptic Transmission genetics
Amino Acid Motifs genetics
Clathrin metabolism
Endocytosis physiology
Monomeric Clathrin Assembly Proteins
Nerve Tissue Proteins metabolism
Phosphoproteins metabolism
Synaptic Vesicles metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1529-2401
- Volume :
- 20
- Issue :
- 23
- Database :
- MEDLINE
- Journal :
- The Journal of neuroscience : the official journal of the Society for Neuroscience
- Publication Type :
- Academic Journal
- Accession number :
- 11102472