Fig. 3.

MinDE spatiotemporally position a lipid-anchored protein resulting in large-scale concentration gradients. a MinDE self-organization spatiotemporally regulates lipid-anchored streptavidin. Representative time series of MinDE self-organization on a SLB with Biotinyl-CAP-PE-bound streptavidin (1 µM MinD, 1 µM MinE, streptavidin-Alexa647). ATP is added at t = 0 s to start self-organization. Scale bars: 50 µm. b Schematic of the experimental setup. Tetrameric streptavidin is anchored to the SLB by binding two to three Biotinyl-CAP-PE lipids and MinDE and ATP are added. c Kymograph of the line selections shown in a. Scale bars: 50 µm and 10 min. d MinDE self-organization leads to large-scale concentration gradients of streptavidin. Representative images of streptavidin distribution in MinDE spirals after >1 h of MinDE self-organization on SLBs. Fluorescence intensity line plots of EGFP-MinD and streptavidin distribution of selections shown in the respective images. Scale bars: 50 µm. e Large-scale streptavidin gradient formation by MinDE is reversible. Representative images and kymograph (1) of a running MinDE assay in the presence of anchored streptavidin. Addition of sodium orthovanadate (Na₃VO₄) leads to MinDE detachment which in turn leads to homogenization of streptavidin fluorescence on the membrane. Fluorescence intensity of streptavidin (cyan) and EGFP-MinD (green) is plotted over the duration of the time-lapse in the center (2) and at the rim of the MinDE spiral (3). Scale bars: 50 µm and 300 s. All experiments were performed independently three or more times under identical conditions