Figure 1

Syntaxin peptide can be immobilized on solid support and can form the SNARE complex. (A) Schematic showing the immobilization strategy. A fusion containing protein of interest (e.g. enzyme) and brevin can be produced by recombinant means. SNAP25, a two-helical protein, can link brevin and syntaxin into a stable tetra-helical bundle. In the sequence of syntaxin peptide, the fluorescein group (FITC) is linked to the N-terminal glutamate via aminohexaenoic acid (Ahx). The native lysine 204 is replaced by arginine (black) allowing cross-linking to solid support only through the newly introduced C-terminal lysines. (B) Image of syntaxin fluorescent beads obtained on a confocal microscope. Scale bar is 50 μM. (C) SDS-PAGE Coomassie-stained gel showing that SNAP25, brevin and the syntaxin peptide assemble into a SDS-resistant complex in a 30 min reaction. Molecular weights are indicated on the left.