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. 2017 Aug 17;7:8648. doi: 10.1038/s41598-017-08917-8

Figure 5.

Figure 5

Disaggregation reaction of aggregated EYFP tethered by a chaperone. (a) Schematic drawing of the construction of YFP-TB and YFP-TK_NBD-B are shown. (b) YFP-TB (left panel), YFP-TK_NBD-B (right panel), and their mutants (6.0  μM monomer) were heat treated at 80 °C for 15 min and the EYFP portions of these proteins were aggregated. The heat-treated fusion proteins were diluted to 0.3 μM monomer, incubated at 55 °C, and the fluorescence at 527 nm (excitation 513 nm) was monitored. After 2 min incubation, ATP (final concentration was 5 mM) was added and the fluorescence monitoring was continued. Fluorescence intensities are shown as a percentage of that prior to heat aggregation. (c) Initial rates of the recovery of the fluorescence were calculated from the measurements shown in (b). Error bars represent standard deviations of three or more independent measurements. WT, wild-type.