STIMULATION OF NSF ATPASE ACTIVITY DURING T-SNARE PRIMING

N-Ethylmaleimide-sensitive factor (NSF) plays a key role in vesicular traffic by disassembling and priming SNARE proteins for their function in docking and fusion. We demonstrate that the ATPase activity of NSF is activated by alpha-soluble NSF attachment protein (alpha-SNAP) in a complex with syntaxin 1A. In addition, me show that a construct cons isting of the H3 domain of syntaxin 1A (GST-synt(195-263), which does not support NSF disassembly in the presence of MgATP gave a larger sti mulation. NSF ATPase activation was specific and did not occur using m utant alpha-SNAPs unable to bind GST-synt or with mutated C-termini. W e suggest that activation of NSF ATPase activity in the SNARE complex may be essential to allow SNARE priming. (C) 1998 Federation of Europe an Biochemical Societies.