FIGURE 1.

Refolding kinetics of GFP mediated by single ring variant SR1 and GroES. A, refolding kinetics of acid-denatured GFP monitored by fluorescence at 509 nm in the absence of SR1 and GroES (black), in the presence of 0.8 μm SR1 and 1.6 μm GroES (red), and in the presence of 2.0 μm SR1ΔC and 4.0 μm GroES (blue). In the red and blue traces, 2 mm ATP was added 200 s after dilution of acid-denatured GFP by the refolding buffer, which contained SR1 (SR1ΔC) and GroES. B, size exclusion chromatography of refolded GFP in the presence of SR1 (red) or SR1ΔC (blue) and GroES was monitored by fluorescence at 509 nm. After monitoring the refolding kinetics by fluorescence (25 min after the addition of ATP), an aliquot of the mixture was subjected to chromatography. The yield of encapsulation without the contribution of spontaneous refolding was calculated by Equation 1. C, the stability of the SR1ΔC·ES/GFP ternary complex. Twenty-five (blue) or 145 min (green) after the addition of ATP to trigger refolding, the mixture was subjected to size exclusion chromatography. a.u., arbitrary units.