FIG. 7.
Interaction of MinD mutants with phospholipid vesicles, MinC, and MinE. (A) AAA (4 μM) was incubated with phospholipid vesicles (400 μg/ml) in ATPase buffer in the presence of 1 mM ADP or ATP. Either no additions were made or MalE-MinC116-231 (4 μM), MinE (4 μM), or both (4 μM) were added, and the samples were incubated at room temperature for 10 min. The samples were centrifuged to sediment the vesicles, and the pellets were analyzed by SDS-PAGE. (B) D152A (4 μM) was incubated with phospholipid vesicles (400 μg/ml) in ATPase buffer in the presence of 1 mM ADP or ATP or no nucleotide. Either no additions were made or MalE-MinC116-231 (4 μM), MinE (4 μM) or both (4 μM each) were added, and the amount of proteins bound to vesicles was determined after centrifugation and analysis of the pellets by SDS-PAGE.