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. 2012 Apr 27;2:386. doi: 10.1038/srep00386

Figure 2. DM promotes release of peptides from the S form of the complex only in the presence of an exchange peptide.

Figure 2

(A) FP analysis of peptide release was carried out in three phases. In phase I, purified FAM-HASG/DR1 complexes were co-incubated with 3 fold excess DM and in the absence of exchange peptide until reaction steady state was reached. Then DM was removed on an anti-FLAG column and free peptide by filtration though centrifugal filter device. Re-purified complexes were then co-incubated with DM (phase II) for 4 hours in the absence of exchange peptide, followed by addition of unlabeled HASG in equimolar amount as compared to complex concentration (phase III). Data are plotted as the fraction of complex remaining relative to t = 0. Reactions were performed in triplicate, and data points represent mean ± SD of three independent experiments. (B) – (C) SDS-PAGE gel analysis (B) and relative quantitation (C) of samples at the beginning and end of phase I (lane 1 and 2), and at the beginning and end of phase II (lane 3 and 4). L, DM labile conformer; S, DM-stabile conformer.