Extended Data Fig. 1: Biophysical data from LOCKR design.
a, Size-exclusion chromatography for the monomer, truncation and LOCKR designs on Superdex 75. Peaks indicated by vertical dashed lines represent monomeric protein used in downstream characterization and functional assays. Size-exclusion chromatography was repeated three times with similar results. b, Circular dichroism spectroscopy to determine protein stability upon heating and treatment with the chemical denaturant guanidinium chloride. Top, full wavescan at 25 °C (blue), 75 °C (orange) and 95 °C (red), then cooled to 25 °C (cyan). Middle, guanidinium chloride melts (also shown overlapped in Fig. 1d). Bottom, fraction of folded protein was converted to the equilibrium constant, and then to the conformational stability for protein unfolding (ΔGunfolding) value. The linear unfolding region, marked by vertical lines in the middle panels, was fit to determine the ΔGfolding for each design. The experiment was repeated four times with similar results. c, SAXS spectra (black; referenced in Fig. 1e) fit to the Rosetta design models (red) using FoXS with χ-values referenced in the top right.
