Identification of Residues Important for the Activity of Haloferax volcanii AglD, a Component of the Archaeal N-Glycosylation Pathway

Figure 3

Hfx. volcanii AglD Asp110 and Asp112 residues likely participate in the GT2 DXD motif involved in the catalytic activity of the enzyme. Site-directed mutagenesis was performed to generate CBD-AglD containing mutations of Asp110 (left column) or Asp112 (right column), as listed on the left of each panel. For each mutant, the upper and lower panels, respectively, show the Coomassie- and PAS-stained S-layer glycoprotein from the background strain (lanes 1 and 5), from the aglD deletion strain (lane 2), from the aglD deletion strain complemented with a plasmid encoding CBD-AglD (lane 3), or from the aglD deletion strain complemented with a plasmid encoding CBD fused to mutated AglD (lane 4).