Figure 5.

Chemical cross-linking of rat brain membrane reveals complexes containing syntaxin1A and nSec1, but not VAMP2 or SNAP-25. Rat brain membrane detergent extracts were incubated with or without DSS (1 mM) for 2 h on ice. Lysates (20 μg/lane) were analyzed by Western blot with the antisyntaxin1A, anti-nSec1, anti–SNAP-25, or anti-VAMP2 antibodies as indicated. Asterisk-labeled complexes are described in Results. There are also two major unidentified cross-linked bands with molecular weights ranging from 45–60 kD recognized by anti–SNAP-25 antibody. There is one major 56-kD cross-linked band recognized by anti-VAMP2 antibody corresponding to synaptophysin/VAMP2 complex as previously demonstrated ( Calakos and Scheller 1994). Molecular mass markers are indicated on the left in kD.