Figure 6.

Gel filtration of unfolded GFP-ClpA complex. A 160-pmol sample of acid-denatured [³H]GFP (A) or native [³H]GFP (B) was incubated with 200 pmol of ClpA in 100-μl reaction mixtures containing buffer A and 1 mM ATP[γS] for 5 min at 24°C. The mixtures were then applied to a 0.7 × 15 cm column of Sephacryl S200 HR as described in the legend of Fig. 3. Initial fluorescence (blue circles) was measured in the fractions. In A 10 mM ATP was then added to each fraction, and after 25 min at 25°C, fluorescence was measured again (red triangles). Radioactivity (open black squares) was measured in aliquots of the fractions.