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. Author manuscript; available in PMC: 2012 Sep 1.
Published in final edited form as: Bioorg Med Chem Lett. 2011 May 24;21(17):4989–4992. doi: 10.1016/j.bmcl.2011.05.038

Figure 3. Analysis of fucosylated glycoproteins and the fucose metabolic pathways in Bacteroidales species.

Figure 3

(a) FucAl is incorporated into Bacteroidales glycoproteins. B. fragilis (lane 1–3) and P. distasonis (lane 4–6) were cultured in the presence (lane 1, 2, 4, 5) or absence (lane 3, 6) of FucAl (200 μM), lysed, treated with (lane 2, 5) or without (lane 1, 3, 4, 6) protease K (100 μg/mL), reacted with biotin-azide via CuAAC and then probed with an α-biotin antibody. (b) P. distasonis is efficiently labeled using low concentrations of FucAl. P. distasonis was cultured in the presence of 200 μM (lane 1), 50 μM (lane 2) and 20 μM (lane 3) FucAl, lysed, reacted with biotin-azide via CuAAC and then probed with an α-biotin antibody. (c) Loss of the B. fragilis fucose isomerase pathway results in greater FucAl incorporation. P. distasonis, B. fragilis and B. fragilis ΔfucI were cultured in the presence of FucAl (200 μM), lysed, reacted with biotin-azide via CuAAC and then probed with an α-biotin antibody. Equal protein loading was confirmed by Coomassie blue staining (data not shown). (d) FKP expression level is not a major factor in comparative FucAl incorporation. Histogram of the relative ratio of FKP mRNA transcript levels between P. distasonis and B. fragilis. Errors were based on triplicate runs in one experiment.