Figure 5
Expression of PatA/PatB in Lemo21(DE3) strain and functionality of the transporter. (A) Membrane proteins in Lemo21(DE3) and C41(DE3) strains were visualized by 12% SDS-PAGE stained with Coomassie blue. Twenty µg of proteins were loaded in each lane. As a negative control, a mutant of PatA/PatB was used in which the conserved Walker-A lysine was mutated in each subunit (PatA-K367A/PatB-K388A). (B) Hoechst 33342 transport assays with inverted membrane vesicles prepared from the various strains. 50 µg of total membrane proteins were used for each assay. (C) Hoechst 33342 transport rates as measured by initial slopes following GTP addition and calculated from panel B. The data represent the average of triplicates, and error bars indicate the standard deviation.
