Transiently receptive DR1 molecules interact with DM/DO complexes.

<p>Mags.DO5 purified DM/DO was immobilized on anti-His antibody coupled chip surface to a level of 2000–3000 RU. Receptive DR1 molecules were generated from DR1/HA(Y308A) peptide complexes by removing the excess peptide by G-50 size-exclusion spin columns, adding DM and allowing the peptide to dissociate for 20 minutes at 37°C. Immediately after the 20 minute incubation 4 µM receptive DR1 with 2 µM DM was injected over the captured DM/DO at a rate of 1 µL/min and a constant flow-cell temperature of 37°C for 50 minutes (green trace). As a control, prior to the injection of receptive DR1 molecules, 2 µM DM (blue trace), or 4 µM DR1/HA(306–318) (red trace) were injected at the same flow rate and temperature. Data shown are representative of two independent experiments.