t-SNARE protein conformations patterned by the lipid microenvironment

Colin Rickman, Claire N Medine, Alison R Dun, David J Moulton, Ondrej Mandula, Nagaraj D Halemani, Silvio O Rizzoli, Luke H Chamberlain, Rory R Duncan

Research output: Contribution to journalArticlepeer-review

52 Citations (Scopus)

Abstract

The spatial distribution of the target (t-)SNARE proteins (syntaxin and SNAP-25) on the plasma membrane has been extensively characterized. However, the protein conformations and interactions of the two t-SNAREs in situ remain poorly defined. By using super-resolution optical techniques and fluorescence lifetime imaging microscopy, we observed that within the t-SNARE clusters syntaxin and SNAP-25 molecules interact, forming two distinct conformations of the t-SNARE binary intermediate. These are spatially segregated on the plasma membrane with each cluster exhibiting predominantly one of the two conformations, representing the two- and three-helical forms previously observed in vitro. We sought to explain why these two t-SNARE intermediate conformations exist in spatially distinct clusters on the plasma membrane. By disrupting plasma membrane lipid order, we found that all of the t-SNARE clusters now adopted a single conformational state corresponding to the three helical t-SNARE intermediates. Together, our results define spatially distinct t-SNARE intermediate states on the plasma membrane and how the conformation adopted can be patterned by the underlying lipid environment.
Original languageEnglish
Pages (from-to)13535-13541
Number of pages7
JournalJournal of Biological Chemistry
Volume285
Issue number18
DOIs
Publication statusPublished - 30 Apr 2010

Keywords

  • animals
  • cell membrane
  • membrane lipids
  • PC12 cells
  • protein structure, quaternary
  • protein structure, secondary
  • Qa-SNARE proteins
  • rats
  • synaptosomal-associated protein 25

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