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. 2018 Dec 12;7:e41771. doi: 10.7554/eLife.41771

Figure 7. Template complex facilitates SNARE assembly.

(A) Representative FECs obtained in the presence of 60 nM SNAP-25B and 2 µM WT Munc18-1 (#1–5 ) or Munc18-1 mutants P335A (#6) or T248G (#7). FECs #1–5 represent consecutive rounds of manipulation of a single Qa-R SNARE conjugate. See also Figure 7—figure supplement 1 and Figure 7—source data 1. (B) Extension-time trajectories at the indicated constant mean forces showing SNARE assembly. Traces a-d were extracted from FEC regions marked with correspondingly labeled red arrows in panel A. Trace f shows rapid template complex transitions without SNAP-25B binding. See also Figure 7—figure supplement 2 and Figure 7—source data 2.

Figure 7—source data 1. MATLAB figure corresponding to Figure 7A (FECs #1–5).
elife-41771-fig7-data1.zip (1.2MB, zip)
DOI: 10.7554/eLife.41771.033
Figure 7—source data 2. MATLAB figure corresponding to Figure 7B.
elife-41771-fig7-data2.zip (158.9KB, zip)
DOI: 10.7554/eLife.41771.034

Figure 7.

Figure 7—figure supplement 1. FECs obtained in the presence of 2 µM Munc18-1 and 60 nM SNAP-25B in the solution.

Figure 7—figure supplement 1.

Red arrows mark events of SNAP-25B binding and SNARE assembly. Note that syntaxin +2 layer mutation I233G/E234G/Y235G significantly weakens CTD zippering.
Figure 7—figure supplement 2. Extension-time trajectories at constant mean forces.

Figure 7—figure supplement 2.

The trajectories exhibit reversible folding and unfolding transitions of the mutant template complexes and irreversible SNAP-25 binding (indicated by red arrows).