Figure 7.

ClpG has high unfolding power. (a) Luciferase-YFP was incubated at 46°C leading to unfolding of the Luciferase moiety and the formation of mixed aggregates comprising misfolded Luciferase and native YFP. YFP fluorescence was monitored during disaggregation reactions as readout for unfolding power of the disaggregases. (b,c) Aggregated Luciferase-YFP was incubated with ClpB, ClpB-K476C or ClpG. Disaggregation reactions with ClpB and ClpB-K476C included the cooperating DnaK system. Disaggregation was monitored by light scattering (b). Sample turbidities at 0 min were set to 100%. Disaggregation rates were determined based on the linear decrease in sample turbidity. Changes in YFP fluorescence were simultaneously recorded (c). Initial YFP fluorescence was set as 100%. YFP unfolding rates were determined based on the linear decrease of YFP fluorescence. Standard deviations are provided (n = 3).