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. 2020 Aug 18;9:e57154. doi: 10.7554/eLife.57154

Figure 8. Effects of phospholipids on C2AB* binding to the SNARE complex.

(A–B) FRET efficiencies observed in titrations of WT C2AB* with solubleSC, cisSC-PC-NDs (containing only PC) or cisSC-PC/PS/PIP2-NDs (containing 15% PS,1% PIP2, 84% PC) labeled with TMR at residue 76 of SNAP-25 in the presence of 1 mM EGTA (A) or 1 mM Ca2+ (B). (C–H) FRET efficiencies in analogous titrations of WT, R398Q/R399Q (RQRQ), R322E/K325E (REKE) or E295A/Y338W (EAYW) C2AB* with solubleSC (C–D), cisSC-PC-NDs (E–F) or cisSC-PC/PS/PIP2-NDs (G–H) in the presence of 1 mM EGTA (C, E, G) or 1 mM Ca2+ (D, F, H). All data were fit with a Hill equation (see Materials and methods).

Figure 8—source data 1. Source data for Figure 8.

Figure 8.

Figure 8—figure supplement 1. Effects of phospholipids on C2AB* binding to the SNARE complex.

Figure 8—figure supplement 1.

(A–B) FRET efficiencies observed in titrations of WT C2AB* with solubleSC or SNARE complex containing the SNARE motif of syntaxin-1 (residues 191–253) (mcc) instead of syntaxin-1 (2–253), both labeled with TMR at residue 76 of SNAP-25, in the presence of 1 mM EGTA (A) or 1 mM Ca2+ (B). (C–D) FRET efficiencies in analogous titrations of WT, R398Q/R399Q (RQRQ), R322E/K325E (REKE) or E295A/Y338W (EAYW) C2AB* with mcc in the presence of 1 mM EGTA (C) or 1 mM Ca2+ (D). All data were fit with a Hill equation (see Materials and methods).
Figure 8—figure supplement 1—source data 1. Source data for Figure 8—figure supplement 1.