Figure 3.

Western immunoblot of the phosphorylation state of His ₆ -Wzd and GST-Wze proteins expressed in E. coli. Tyrosine phosphorylation of His₆-Wzd and GST-Wze was detected by using Western immunoblotting with a mouse monoclonal antibody. Coomassie stained 12% SDS-PAGE gel of His₆-Wzd expressed in E. coli BL21(DE3) (A) and C41(DE3) (B): incubated with 10 mM ATP (Lane 1) and without ATP (Lane 2). (C) Detection of tyrosine phosphorylated His₆-Wzd expressed in E. coli BL21(DE3) and incubated with 10 mM ATP (Lane 1) and without ATP (Lane 2). (D) Absence of detection of tyrosine phosphorylated His₆-Wzd expressed in E. coli C41(DE3) and incubated with 10 mM ATP (Lane 1) and without ATP (Lane 2). (E) Detection of tyrosine phosphorylated GST-Wze expressed in E. coli C41(DE3) incubated with 10 mM ATP (Lane 1) and without ATP (Lane 2).