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

Ilaria Monaldi, Massimo Vassalli, Angela Bachi, Silvia Giovedi, Enrico Millo, Flavia Valtorta, Roberto Raiteri, Fabio Benfenati, Anna Fassio

Research output: Contribution to journalArticle

Abstract

Synapsins are abundant SV (synaptic vesicle)-associated phosphoproteins that regulate synapse formation and function. The highly conserved C-terminal domain E was shown to contribute to several synapsin functions, ranging from formation of the SV reserve pool to 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 (Förster 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 on the size and integrity of SV pools are contributed by the regulation of synapsin dimerization and SV clustering.

Original languageEnglish
Pages (from-to)55-64
Number of pages10
JournalBiochemical Journal
Volume426
Issue number1
DOIs
Publication statusPublished - Feb 15 2010

Fingerprint

Synapsins
Dimerization
Cluster Analysis
Phospholipids
Synaptic Vesicles
Liposomes
Oligomerization
Assays
Affinity chromatography
Peptides
Atomic Force Microscopy
Phosphoproteins
Energy Transfer
Physiology
Exocytosis
Affinity Chromatography
Synapses
Energy transfer
Atomic force microscopy

Keywords

  • Atomic force microscopy (AFM)
  • Exocytosis
  • Förster resonance energy transfer (FRET)
  • Liposome
  • Synapsin
  • Synaptic vesicle

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

The highly conserved synapsin domain E mediates synapsin dimerization and phospholipid vesicle clustering. / Monaldi, Ilaria; Vassalli, Massimo; Bachi, Angela; Giovedi, Silvia; Millo, Enrico; Valtorta, Flavia; Raiteri, Roberto; Benfenati, Fabio; Fassio, Anna.

In: Biochemical Journal, Vol. 426, No. 1, 15.02.2010, p. 55-64.

Research output: Contribution to journalArticle

Monaldi, I, Vassalli, M, Bachi, A, Giovedi, S, Millo, E, Valtorta, F, Raiteri, R, Benfenati, F & Fassio, A 2010, 'The highly conserved synapsin domain E mediates synapsin dimerization and phospholipid vesicle clustering', Biochemical Journal, vol. 426, no. 1, pp. 55-64. https://doi.org/10.1042/BJ20090762
Monaldi, Ilaria ; Vassalli, Massimo ; Bachi, Angela ; Giovedi, Silvia ; Millo, Enrico ; Valtorta, Flavia ; Raiteri, Roberto ; Benfenati, Fabio ; Fassio, Anna. / The highly conserved synapsin domain E mediates synapsin dimerization and phospholipid vesicle clustering. In: Biochemical Journal. 2010 ; Vol. 426, No. 1. pp. 55-64.
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