Department of Anesthesiology

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A distributed set of interactions controls mu2 functionality in the role of AP-2 as a sorting adaptor in synaptic vesicle endocytosis.

TitleA distributed set of interactions controls mu2 functionality in the role of AP-2 as a sorting adaptor in synaptic vesicle endocytosis.
Publication TypeJournal Article
Year of Publication2009
AuthorsKim SHyun, Ryan TA
JournalJ Biol Chem
Volume284
Issue47
Pagination32803-12
Date Published2009 Nov 20
ISSN1083-351X
KeywordsAction Potentials, Adaptor Protein Complex 2, Adaptor Protein Complex mu Subunits, Animals, Clathrin, Clathrin-Coated Vesicles, Endocytosis, Exocytosis, Hippocampus, Hydrogen-Ion Concentration, Kinetics, Mutation, Neurons, Protein Structure, Tertiary, Rats, Synaptic Vesicles
Abstract

The mechanisms of how, following exocytosis, the approximately nine types of synaptic vesicle (SV) transmembrane proteins are accurately resorted to form SVs are poorly understood. The time course of SV endocytosis is very sensitive to perturbations in clathrin and dynamin, supporting the model that SV endocytosis occurs through a clathrin-mediated pathway. We recently demonstrated that removal of the clathrin adaptor protein AP-2, the key protein thought to coordinate cargo selection into clathrin-coated pits, results in a significant impairment in endocytosis kinetics. Endocytosis, however, still proceeds in the absence of AP-2, bringing into question the role of AP-2 in cargo sorting in this process. Using quantitative endocytosis assays at nerve terminals, we examined how endocytosis depends on the integrity of mu2 function. Our experiments indicate that no single perturbation in mu2 prevents restoration of endocytic function when mutated mu2 replaces native mu2, whereas introduction of multiple distributed mutations significantly impairs endocytosis. We also examined whether the presence of AP-2 is important for the functionality of the previously identified endocytic motif in an SV cargo protein, the dileucine motif in vGlut-1. These data show that while mutations in the dileucine motif slow the retrieval of vGlut-1, they only do so in the presence of AP-2. These data thus indicate that AP-2 plays a role in cargo selection but that no single aspect of mu2 function is critical, implying that a more distributed network of interactions supports AP-2 function in SV endocytosis.

DOI10.1074/jbc.M109.039149
Alternate JournalJ. Biol. Chem.
PubMed ID19762466
PubMed Central IDPMC2781697
Grant List36942 / / PHS HHS / United States
R01 NS036942 / NS / NINDS NIH HHS / United States