Fig. 2

Binding assays with inside-out vesicles (IOVs). a Expression and purification of recombinant SURFIN ^WRD2_4.2 -His. The Coomassie Brilliant Blue staining (left) and Western blot analysis (middle) of the total E. coli lysates of the uninduced (Pre), induced, and purified culture of recombinant SURFIN ^WRD2_4.2 -His (calculated MW, 46.5 kDa). Right panel shows a Western blot for the bound, washed, and eluted fractions of the His tag alone (20.4 kDa). The bands at the predicted size for the recombinant protein were marked with arrowheads. b A saturation binding curve using ELISA-based protein interaction analysis. The dissociation constant (K _d) was determined to be 0.58 ± 0.02 μM. c Scatchard plot of SURFIN ^WRD2_4.2 -His and IOVs interaction. B _max = 0.44 ± 0.03 μM