Figure 5.

The N417ins mutation increases membrane tethering efficiency of the ATL1 catalytic-core fragment.A, select tethering reactions at increasing protein concentrations (between 0.5 and 1.5 μM protein) for ATL1 WT (blue) and N417ins (maroon). Each point is the average of two biological and three technical replicates, with gray error bars for the SD. B, initial tethering rates (min⁻¹) were calculated for each ATL1 concentration, displayed as the mean and SD across replicates. The inset (right panel) shows concentrations used to calculate values plotted in C (reactions with 0.75–1.5 μM protein). C, specific tethering rates (min⁻¹ μM⁻¹) for each construct’s technical and biological replicates. An unpaired t test was used to determine the significance between the WT and mutant rates (p = 0.0237). D, images (top views of the individual wells of a 96-well plate; well diameter = 5 mm) of tethering reactions after 45 min ± GTP at 1.5 μM.