Botulinum E toxin light chain does not cleave SNAP-23 and only partially impairs insulin stimulation of GLUT4 translocation in 3T3-L1 cells

S. Lance MacAulay, Shane Rea, Keith H. Gough, Colin W. Ward, David E. James

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

The stimulation of glucose uptake into fat and muscle by insulin results predominantly from the translocation of the glucose transporter, GLUT4, from an intracellular vesicle pool to the cell surface. Homologues of several key proteins known to be involved in the process of synaptic vesicle fusion have been identified on GLUT4 vesicles, including VAMPS and cellubrevin. Syntaxin 4, SNAP-23 and/or SNAP-25 are also implicated in this process. Bacterial toxins that specifically cleave these proteins have been utilised to assess their involvement in cell function. We aimed to distinguish which of the SNAP isoforms are specifically involved in GLUT 4 translocation. Here we show that both human (h) and mouse (m) SNAP-23, unlike SNAP-25, are not substrates for Botulinum E toxin light chain (BoNT/E). Furthermore, we demonstrate that microinjection of differentiated 3T3-L1 cells with BoNT/E inhibited insulin stimulation of GLUT4 translocation only slightly, 27%, whereas tetanus toxin light chain, that cleaves VAMP2, inhibited insulin stimulation of GLUT4 translocation by 80%. These studies therefore do not support a major role for SNAP 25 in insulin stimulation of GLUT4 translocation and place SNAP-23 as a prime candidate for a role in this process.

Original languageEnglish (US)
Pages (from-to)388-393
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume237
Issue number2
DOIs
StatePublished - Aug 18 1997

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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