The highly conserved synapsin domain E mediates synapsin dimerization and phospholipid vesicle clustering

Synapsins are abundant SV (synaptic vesicle)-associated phosphoproteins that regulate synapse formation and function. Title highly conserved C-terminal domain E wits shown to contribute to several synapsin functions. ranging from formation of the SV reserve pool to ne regulation of the kinetics of exocytosis and SV cycling. although the Molecular mechanisms underlying these effects are unknown. In the present study, We used a synthetic 25-mer peptide encompassing the most conserved region of domain E (Pep-E) to analyse the role of domain E in regulating the interactions between synapsin I and liposomes mimicking the phospholipid composition of SVs (SV-liposomes) and other pre-synaptic protein partners. In affinity-chromatography and cross-linking assays, Pep-E bound to endogenous and purified exogenous synapsin I and strongly inhibited synapsin dimerization, indicating a role in synapsin oligomerization. Consistently, Pep-E (but not its scrambled version) Counteracted the ability of holo-synapsin I to bind and coat phospholipid membranes, as analysed by AFM (atomic force microscopy) topographical scanning, and significantly decreased the clustering of SV-liposomes induced by holo-synapsin I in FRET (Forster resonance energy transfer) assays, suggesting a causal relationship between synapsin oligomerization and vesicle clustering. Either. Pep-E or a peptide derived from domain C Was necessary and sufficient to inhibit both dimerization and vesicle clustering. indicating the participation of both domains in these activities of synapsin I. The results provide a molecular explanation for the effects of domain E in nerve terminal physiology and suggest that its effects oil the size and integrity of SV pools are Contributed by the regulation of synapsin dimerization and SV clustering.