Fig. 4
Glycosylation of ApxIVAC1, ApxIVAC2 and ApxIIAD3 (Ca2+-binding domain). a- Schematic representation of ApxIVAC1, ApxIVAC2 and ApxIAD3. Each peptide was engineered to include N- and C-terminal NAT sequons. ApxIAD3 was further modified to include an internal NDT motif. b- The toxin fragments were expressed in E. coli with and without chromosomally encoded ngt and agt. The fragments were purified by Ni-NTA affinity chromatography, resolved by SDS-PAGE and detected by coomassie staining. c- The following Ni-NTA purified fragments were resolved by SDS-PAGE and PAS stained: Lane 1: ApxIA (no NX(S/T) sequons); Lane 2: ApxIA (G71 T), no ngt and agt Lane 3: ApxIA (G71 T) plus chromosomally encoded ngt and agt; Lane 4: ApxIIA, no ngt and agt; Lane 5: ApxIIA plus chromosomally encoded ngt and agt; Lane 6: AtaC, plasmid-encoded ngt and agt