Figure 6.

Degradation of GFP-SsrA by ClpX. (A) Identical solutions with 1 μM each of GFP-SsrA, ClpX, and ClpP in degradation buffer with 10 mM ATP were incubated at 37°C. For one, the fluorescence was monitored, and, for the other, aliquots were withdrawn at various times and prepared for SDS/PAGE. The GFP-SsrA remaining was detected by silver staining (Inset). Numbers above lanes are the times of incubation with ClpXP. (B) GFP-SsrA was subjected to two different treatments, and the kinetics of degradation in the presence of 0.5 μM ClpP and 5 mM ATP was measured. In the first, GFP-SsrA was unfolded by 1 μM ClpX and diluted 10-fold into degradation buffer (●). In the second, GFP-SsrA was incubated without ClpX and then diluted into degradation buffer containing 0.1 μM ClpX in addition to the other components (○).