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. 1998 Jan 26;140(2):431–446. doi: 10.1083/jcb.140.2.431

Figure 6.

Figure 6

Figure 6

Figure 6

Effect of glycosylation on the intrinsic CD44Rg affinity for HA. Affinity capillary electrophoresis in the presence of the indicated concentrations of HA60 of (A) wt CHO–derived CD44Rg, (B) BSA, or (C–F) ldl-D–derived CD44Rg synthesized in the presence of (C) no saccharide addition, (D) Gal only, (E) GalNAc only, or (F) Gal + GalNAc. The narrow peak due to MO, a neutral marker included as an internal control, is labeled. The broader unlabeled peak corresponds to CD44Rg. The inverted peak at the migration time for HA60 is due to the absence of HA60 in the sample. Mobility values for CD44Rg and HA60 were calculated from migration times relative to MO. The mobility of CD44Rg in the presence of ligand is reported as a relative mobility (R M), where R M is 0.0 at the mobility of the uncomplexed CD44Rg and R M is 1.0 at the mobility of HA60. The migration times corresponding to R M of 0.0 and 0.4, indicated in each panel for the top electrophoretogram, were calculated using the mobility of CD44Rg from the CE runs at 0 μM HA60 and the migration times for MO and HA60 from the same electrophoretogram. The relative mobilities of CD44Rg from A–F are plotted in G. Displayed curves were fit to the equation R M = R M (max) * [HA60]/(K d * [HA60]) where R M is the relative mobility, R M (max) is the maximum relative mobility (mobility of CD44Rg at saturating concentrations of HA60), K d is the dissociation constant, and [HA60] is the concentration of HA 60-mer. The determined values were R M (max), 0.54 and K d, 5 and 40 μM.