Figure 5.

Dissociation of wt and DR1_βG86Y complexes by DM in real time. Peptide–DR1 surfaces were generated as in the legend to Fig. 4, and 9 μM DM was passed through four different peptide–DR1 complex surfaces. For better comparison, all surfaces indicating different RU values were given zero values as the initiation point. DR1_βG86Y–HA surface was used as a negative control, as binding was negligible (Fig. 4). The negative sensograms are due to lower salt concentration in DM-containing buffer, and perhaps lack of Tween 20 that was present in the running buffer. Shown are wtDR1–HA_Anchorless (red), wtDR1–HA (blue), DR1_βG86Y–HA_Anchorless (red), and the control mock DR1_βG86Y–HA (blue). In the presence of DM, dissociation rates for the wtDR1–HA_Anchorless were estimated to be 3.8 × 10⁻³ s⁻¹ (t _1/2 ∼ 3.0 min); for wtDR1–HA, 5.6 × 10⁻⁷ s⁻¹ (t _1/2 ∼ 344 h); and for the DR1_βG86Y–HA_Anchorless, 8.5 × 10⁻⁵ s⁻¹ (t _1/2 ∼ 2.25 h). Inset, a replot of data after subtraction of the mock DR1_βG86Y–HA control surface to provide an easy evaluation of the dissociation rates of the complexes in the presence of DM. One out of two experiments is shown.