Alternative Computational Protocols for Supercharging Protein Surfaces for Reversible Unfolding and Retention of Stability
A) Total fluorescence values indicate the level of expression of correctly folded GFP. The low- to mid-charge negative variants expressed well, but mid- to high-charge variants expressed significantly worse that sfWT. B) Absorbance ratios indicate the relative amount of correctly folded GFP. Absorbance by the chromophore at 495 nm indicates correctly folded GFP, and absorbance at 280 nm indicates the total amount of GFP. Low- to mid-charge variants are well folded (before thermal challenge), while high-charge variants are not well folded.