The N-terminal 31 amino acids of ATL1 are critical for interaction with spastin and spastin-mediated enhancement of ATL1-mediated liposome fusion. (A) Wild-type ATL1, ATL1-Δ(2–31), or ATL1-Δ(521–558) was reconstituted into liposomes with or without M1-spastin. Donor and acceptor liposomes were then mixed and incubated in the presence of GTP. Fusion is represented as the percentage of total NBD fluorescence. Data represent the means ± SEM (error bars; n = 3). n refers to the number of independent experiments. *P < 0.05, two-tailed unpaired Student’s t test. (B) HEK293T cells were transfected with genes encoding EGFP-conjugated ATL1 (EGFP-ATL1) alone, EGFP-ATL1 and myc-tagged Spastin (myc-Spastin), EGFP-ATL1-Δ(2–31) and myc-Spastin, or EGFP-ATL1-Δ(521–558) and myc-Spastin, and the physical interaction between M1-spastin and wild-type or mutant ATL1 was analyzed by co-immunoprecipitation. Source data are available for this figure: SourceData F5.
Figure 5.

The N-terminal 31 amino acids of ATL1 are critical for interaction with spastin and spastin-mediated enhancement of ATL1-mediated liposome fusion. (A) Wild-type ATL1, ATL1-Δ(2–31), or ATL1-Δ(521–558) was reconstituted into liposomes with or without M1-spastin. Donor and acceptor liposomes were then mixed and incubated in the presence of GTP. Fusion is represented as the percentage of total NBD fluorescence. Data represent the means ± SEM (error bars; n = 3). n refers to the number of independent experiments. *P < 0.05, two-tailed unpaired Student’s t test. (B) HEK293T cells were transfected with genes encoding EGFP-conjugated ATL1 (EGFP-ATL1) alone, EGFP-ATL1 and myc-tagged Spastin (myc-Spastin), EGFP-ATL1-Δ(2–31) and myc-Spastin, or EGFP-ATL1-Δ(521–558) and myc-Spastin, and the physical interaction between M1-spastin and wild-type or mutant ATL1 was analyzed by co-immunoprecipitation. Source data are available for this figure: SourceData F5.

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