FtsZ-I374V does not bind MinCC/MinD in vitro. The interaction between FtsZ and MinCC/MinD was determined by a sedimentation assay in which the recruitment of FtsZ polymers to a phospholipid vesicle-associated MinCC/MinD complex was assessed. Reactions containing mutilamellar phospholipid vesicles (MLV, 400 µg/mL), MinD (6µM), WT FtsZ (A) or FtsZ-I374V (B) at 6 µM, GMPCPP or GDP (200 µM) with (lane 3 to 6) or without (lane 1 and 2) MalE-MinCC (6 µM) were incubated at room temperature for 5 min in ATPase buffer (25 mM Tris-HCl [pH 7.5], 50mM KCl, and 5mM MgCl2). ATP or ADP (1 mM) were then added to the reactions and incubated for another 5 min. The reactions were then centrifuged at 10,000×g for 2 min. The pellets were solubilized and analyzed by SDS-PAGE.