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. 2009 Jul 15;106(30):12289–12294. doi: 10.1073/pnas.0905744106

Fig. 3.

Fig. 3.

Kinetics of unfolding of unlabeled and TNB-labeled proteins as monitored by FRET. (A) Cys97-TNB; (B) Cys29-TNB. In A and B, the changes in FRET efficiency during unfolding in 4 M GdnHCl (dark red continuous curve) and 6 M GdnHCl (dark green continuous curve) are shown, and the dashed black line shows the FRET efficiency estimated in the native protein. The data in A and B were converted into D–A distances by using Eq. S2, and the distances are shown for (C) Cys97-TNB and (D) Cys29-TNB at 4 M GdnHCl (dark red continuous curve) and 6 M GdnHCl (dark green continuous curve). The black dashed line shows the D–A distance estimated in the native protein. Identical kinetics of unfolding were seen at both 15 μM and 50 μM protein concentrations, indicating the absence of any transient aggregation during the unfolding reaction.