Figure 3. Non-canonical SNARE complexes support fusion when their surface density is increased.
(A) Standard fusion reactions containing vacuoles from wild type strains (black bars) or SNARE overexpressing strains (gray bars) were blocked at the priming stage with 85 μg/ml anti-Sec17p antibody for 15 minutes. Recombinant GST-Vam7p (400 nM, black bars) or Vam7pQ283R (gray bars) was added to bypass the priming block. Error bars represent SEM (n=3). (B) GST-Vam7p protein complexes were isolated as described in the Materials and Methods section and examined by immunoblotting for the indicated proteins.