Crystal structure and biophysical properties of a complex between the N-terminal SNARE region of SNAP25 and syntaxin 1a

J Biol Chem. 2001 Nov 2;276(44):41301-9. doi: 10.1074/jbc.M106853200. Epub 2001 Aug 30.

Abstract

SNARE proteins are required for intracellular membrane fusion. In the neuron, the plasma membrane SNAREs syntaxin 1a and SNAP25 bind to VAMP2 found on neurotransmitter-containing vesicles. These three proteins contain "SNARE regions" that mediate their association into stable tetrameric coiled-coil structures. Syntaxin 1a contributes one such region, designated H3, and SNAP25 contributes two SNARE regions to the fusogenic complex with VAMP2. Syntaxin 1a H3 (syn1aH3) and SNAP25 can form a stable assembly, which can then be bound by VAMP2 to form the full SNARE complex. Here we show that syn1aH3 can also form a stable but kinetically trapped complex with the N-terminal SNARE region of SNAP25 (S25N). The crystal structure of this complex reveals an extended parallel four-helix bundle similar to that of the core SNARE and the syn1aH3-SNAP25 complexes. The inherent ability of syn1aH3 and S25N to associate stably in vitro implies that the intracellular fusion machinery must prevent formation of, or remove, any non-productive complexes. Comparison with the syn1aH3-SNAP25 complex suggests that the linkage of the N- and C-terminal SNAP25 SNARE regions is kinetically advantageous in preventing formation of the non-productive syn1aH3-S25N complex. We also demonstrate that the syn1aH3-S25N complex can be disassembled by alpha-SNAP and N-ethylmaleimide-sensitive factor.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antigens, Surface / chemistry*
  • Carrier Proteins / chemistry
  • Cells, Cultured
  • Crystallography, X-Ray
  • Kinetics
  • Membrane Proteins / chemistry*
  • Models, Molecular
  • N-Ethylmaleimide-Sensitive Proteins
  • Nerve Tissue Proteins / chemistry*
  • Protein Conformation
  • Rats
  • SNARE Proteins
  • Synaptosomal-Associated Protein 25
  • Syntaxin 1
  • Vesicular Transport Proteins*

Substances

  • Antigens, Surface
  • Carrier Proteins
  • Membrane Proteins
  • Nerve Tissue Proteins
  • SNARE Proteins
  • Snap25 protein, rat
  • Stx1a protein, rat
  • Synaptosomal-Associated Protein 25
  • Syntaxin 1
  • Vesicular Transport Proteins
  • N-Ethylmaleimide-Sensitive Proteins
  • Nsf protein, rat