FIGURE 1:

Purified DDB complex demonstrates processive, diffusive, and stuck behaviors. (A) Schematic of DDB purification using BicD2-coated Strep-Tactin beads to pull dynein/dynactin from brain lysate. DDB was then eluted off of the bead. (B) SDS–PAGE gel of recombinant BicD2 and final purified DDB complex showing dominant bands of dynein heavy chain (DHC), dynactin components p150 and p135, and BicD2. (C) Tagging DDB for single-molecule tracking. Biotinylated GFP binding protein (GBP) is used to link C-terminal GFP on BicD2 to streptavidin-coated quantum dots for TIRF experiments or streptavidin-coated 30-nm gold nanoparticles for iSCAT experiments. (D) Kymograph of kinesin-1 (left) and DDB (right) single-molecule motility. DDB displays processive runs (P), diffusive episodes (D), and stuck events (S).