FIG 5.

Enrichment of OmpA-like glycoproteins from P. gingivalis wild-type and mutant strains by WGA column chromatography. Whole-cell lysates from P. gingivalis wild-type and mutant strains were solubilized with 1% DM and applied to a WGA-agarose minicolumn overnight at 4°C and then washed with 10 bed volumes of 10 mM Tris-HCl (pH 7.4) containing 0.15 M NaCl and 0.03% DM. The bound proteins were eluted with GlcNAc in the same buffer and then extensively dialyzed against 10 mM Tris-HCl (pH 7.4) and concentrated. The concentrated proteins were subjected to SDS-PAGE under nonreducing and reducing conditions and stained with CBB (A) and then further analyzed by Western blotting with an anti-Pgm6/7 antibody (B). Lane M, molecular marker. 2ME, 2-mercaptoethanol.