Figure 7
From: Structure and tailspike glycosidase machinery of ORF212 from E. coli O157:H7 phage CBA120 (TSP3)
Active site mutant halo assays. E. coli strain ATCC 700728 was embedded in agarose. Wells (3 mm) were cut out of the agarose and loaded with 10 µL (6 mg/mL) wild-type TSP3 or active site mutants and incubated overnight to visualize tailspike activity. (a) (1) wild-type TSP3; (2) TSP3 E362Q; (3) TSP D383N; (4) TSP3 D426N; (5) TSP3 E362Q:D383N double mutant; (6) TSP3 E362Q:D426N double mutant; (7) TSP3 D383N:D426N double mutant; (8) TSP3 E362Q:D383N:D426N triple mutant; (9) PBS control. (b) Identical image to (a) with a dashed white line to indicate the halo edge to aid in visualization. Quantitation of Fig. 6 is shown in Table 2.
