Anti-synapsin monoclonal antibodies: Epitope mapping and inhibitory effects on phosphorylation and Grb2 binding

Paola Vaccaro, Luciana Dente, Franco Onofri, Adriana Zucconi, Sabrina Martinelli, Flavia Valtorta, Paul Greengard, Gianni Cesareni, Fabio Benfenati

Research output: Contribution to journalArticle

Abstract

The synapsins are a family of major neuron-specific synaptic vesicle-associated phosphoproteins which play important roles in synaptic function. In an effort to identify molecular tools which can be used to perturb the activity of the synapsins in in vitro as well as in vivo experiments, we have localized the epitopes of a panel of monoclonal antibodies (mAbs) raised against synapsins I and II and have characterized their ability to interfere with the interactions of the synapsins with protein kinases, actin and Src homology-3 (SH3) domains. The epitopes of the six mAbs were found to be concentrated in the N-terminal region within domains A and B for the synapsin II-reactive mAbs 19.4, 19.11, 19.51 and 19.21, and in two C-terminal clusters in the proline-rich domains D for synapsin I (mAbs 10.22, 19.51, 19.11 and 19.8) and G for synapsin II (mAb 19.8). The synapsin II-specific mAbs 19.4 and 19.21, whose overlapping epitopes are adjacent to phosphorylation site 1, specifically inhibited synapsin II phosphorylation by endogenous or exogenous cAMP-dependent protein kinase. While all the anti-synapsin I mAbs were unable to affect the interactions of synapsin I both with Ca2+/calmodulin-dependent protein kinase II and with actin monomers and filaments, mAbs 19.8 and 19.51 were found to inhibit the binding of Grb2 SH3 domains to the proline-rich C-terminal region of synapsin I.

Original languageEnglish
Pages (from-to)1-16
Number of pages16
JournalMolecular Brain Research
Volume52
Issue number1
DOIs
Publication statusPublished - Dec 1 1997

Keywords

  • Actin
  • CaM kinase II
  • Epitope mapping
  • Monoclonal antibody
  • Peptide library
  • Protein kinase A
  • SH3 domain
  • Synapsin
  • Synaptic vesicle

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

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    Vaccaro, P., Dente, L., Onofri, F., Zucconi, A., Martinelli, S., Valtorta, F., Greengard, P., Cesareni, G., & Benfenati, F. (1997). Anti-synapsin monoclonal antibodies: Epitope mapping and inhibitory effects on phosphorylation and Grb2 binding. Molecular Brain Research, 52(1), 1-16. https://doi.org/10.1016/S0169-328X(97)00219-2