Figure 5. Synergistic interactions of C2B with membrane-anchored SNARE complexes, PI(4,5)P2 and PS in the presence of 0.1 mM Ca2+.
(A and B) Co-flotation of Syt1 C2B with liposomes in the absence of PI(4,5)P2 (A) and quantification of the results (B). (C and D) Co-flotation of Syt1 C2B with liposomes in the presence of 1% PI(4,5)P2 (C) and quantification of the results (D). Cis-SNARE complexes were reconstituted on liposome via the syntaxin-1 transmembrane domain. Liposomes compositions in A and C contain 65% PC, 20% PE, 15% PS with and without 1% PI(4,5)P2. S, supernatant; P, pellet. Representative gel from one of three independent experiments is shown. Data are processed by Image J (NIH) and presented as the mean ± SD (n = 3), technical replicates.
Figure 5—figure supplement 1. Binding of C2B to PS-containing liposomes in different Ca2+ concentrations.
(A) Co-flotation of C2B with liposomes (65% PC + 20% PE + 15% PS) in the presence of 250 mM KCl and different Ca2+ concentrations as indicated. (B) Quantification of the results in A. Representative gel from one of three independent experiments is shown. Data are processed by Image J (NIH) and presented as the mean ± SD (n = 3), technical replicates.