Synaptotagmin binding to botulinum neurotoxins

Research output: Contribution to journalArticle

Abstract

Botulinum neurotoxins (BoNTs) are exceptionally toxic proteins that cause paralysis, but are also extensively used as treatment for various medical conditions. Most BoNTs bind two receptors on neuronal cells, namely a ganglioside and a protein receptor. Differences in the sequence between the protein receptors from different species can impact the binding affinity and toxicity of the BoNTs. Here we have investigated how BoNT/B, /DC, and /G, all three toxins that utilize Synaptotagmin I and II (Syt-I/II) as their protein receptors, bind to Syt-I and-II from mouse/rat, bovine and human origin by isothermal titration calorimetry analysis. BoNT/G had the highest affinity to human Syt-I and BoNT/DC had the highest affinity for bovine Syt-II. As expected, BoNT/B, /DC and /G showed very low binding to human Syt-II. Furthermore, we carried out saturation difference transfer (STD) and STD-TOCSY NMR experiments that revealed the region of the Syt peptide in direct contact with BoNT/G, which demonstrate that BoNT/G recognizes the Syt peptide in a similar model as in the established BoNT/B-Syt-II complex. Our analyses also revealed that regions outside the Syt peptide's toxin binding region are important for the helicity of the peptide, and therefore the binding affinity.

Details

Authors
  • Markel Martínez-Carranza
  • Pilar Blasco
  • Robert Gustafsson
  • Min Dong
  • Ronnie P-A Berntsson
  • Göran Widmalm
  • Pål Stenmark
Organisations
External organisations
  • Stockholm University
  • Harvard Medical School
  • Boston Children's Hospital
  • Umeå University
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Medicinal Chemistry
Original languageEnglish
JournalBiochemistry
Publication statusE-pub ahead of print - 2019
Publication categoryResearch
Peer-reviewedYes