Skip to main content
. 2017 Sep 19;6:e27396. doi: 10.7554/eLife.27396

Figure 10. HOPS-SM gates Sec17 function.

Figure 10.

(A and B) RPLs bearing Ypt7:GTP and either the Qa and Qb SNAREs, or the R SNARE, were incubated with Sec17 (0 or 600 nM) and Qc-wt (80 and 400 nM in panel A and B, respectively). In panel A, HOPS was present (100 nM). In panel B, the HOPS requirement was bypassed by adding 2% PEG. In control reactions, Qc-wt or Sec17 were omitted. No Sec18 was present under any condition. (C) RPLs bearing Ypt7:GTP and either the Qa and Qb SNAREs, or the R SNARE, were incubated with Qc-3∆ (80 nM), Sec17 or Sec17 FSMS (600 nM), and HOPS (50 nM) or PEG (2%). All soluble components were added together to the vesicles at t = 0. (D) Reactions were set up as in panel C, except that Sec17 was omitted from the initial reaction mix. After a 10 min pre-incubation at 27°C, Sec17 (wt or the FSMS loop mutant) was added at t = 0. The final reagent concentrations were the same as in panel C. For panels A-D, points denote mean plus or minus s.e.m. of 2 (A,B) or 6 (B,C) independent experiments. Lines show nonlinear best-fits of a second-order kinetic model.