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. Author manuscript; available in PMC: 2021 Jan 26.
Published in final edited form as: Cell Rep. 2021 Jan 12;34(2):108611. doi: 10.1016/j.celrep.2020.108611

Figure 3. Split SNARE-Driven Fusion Is Highly Sensitive to Layer Mutations that Impair Vesicle Fusion In Vivo.

Figure 3.

(A) Sequence of VAMP2 CTDs with layer residues numbered and highlighted.

(B) Liposomes harboring split VAMP2 (shown in Figure 1B, right, with or without layer mutations) were directed to fuse with liposomes containing WT t-SNAREs (syntaxin-1 and SNAP-25). The kinetics of the fusion reactions was measured using a FRET-based lipid-mixing assay.

(C) Initial lipid-mixing rates of the fusion reactions shown in (B). Data are presented as mean ± SD (n = 3). The point mutation data are compared with the no mutation data. The p values were calculated using Student’s t test. ***p < 0.001.

(D) Correlation of the effects of VAMP2 layer mutations on split SNARE-driven liposome fusion, SNARE-SM-mediated liposome fusion, and in vivo vesicle fusion. In vivo data are based on published genetic studies (Walter et al., 2010; Yu et al., 2015). +++++, WT levels of in vitro liposome fusion or in vivo vesicle fusion; +, <20% of WT levels of fusion.