Identification of the synaptic vesicle glycoprotein 2 receptor binding site in botulinum neurotoxin A

FEBS Lett. 2014 Apr 2;588(7):1087-93. doi: 10.1016/j.febslet.2014.02.034. Epub 2014 Feb 25.

Abstract

Botulinum neurotoxins (BoNTs) inhibit neurotransmitter release by hydrolysing SNARE proteins. The most important serotype BoNT/A employs the synaptic vesicle glycoprotein 2 (SV2) isoforms A-C as neuronal receptors. Here, we identified their binding site by blocking SV2 interaction using monoclonal antibodies with characterised epitopes within the cell binding domain (HC). The site is located on the backside of the conserved ganglioside binding pocket at the interface of the HCC and HCN subdomains. The dimension of the binding pocket was characterised in detail by site directed mutagenesis allowing the development of potent inhibitors as well as modifying receptor binding properties.

Keywords: Botulinum neurotoxin A; Monoclonal antibody; Neutralisation; Protein receptor binding site; SV2.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Binding Sites
  • Botulinum Toxins, Type A / chemistry*
  • Botulinum Toxins, Type A / genetics
  • Botulinum Toxins, Type A / pharmacology
  • Humans
  • Inhibitory Concentration 50
  • Membrane Glycoproteins / chemistry*
  • Mice
  • Mice, Inbred C57BL
  • Mutagenesis, Site-Directed
  • Nerve Tissue Proteins / chemistry*
  • Neural Conduction / drug effects
  • Neurotoxins / chemistry*
  • Neurotoxins / genetics
  • Neurotoxins / pharmacology
  • Phrenic Nerve / drug effects
  • Phrenic Nerve / physiology
  • Protein Binding
  • Protein Interaction Domains and Motifs

Substances

  • Membrane Glycoproteins
  • Nerve Tissue Proteins
  • Neurotoxins
  • SV2C protein, human
  • Botulinum Toxins, Type A