Extended Data Fig. 3: Computational and biochemical characterization of HMGB1.

(a) Predicted structures of Hmgb1 proteins from AlphaFold2 Protein Structure Database. Colors ranging from blue to orange depict the per-residue measure of local confidence for the model. (b) Left: MSA depth assessment for the sequences for quality assessment of the predicted HMGB1 models. Aligned sequences are colored by sequence identity. Sequence coverage frequency is depicted by a black line. The dotted red line marks the frameshift in the mutant. Right: Disorder analysis of wild type and mutant HMGB1 sequences using AlphaFold2 pLDDT scores (yellow) and Metapredict scores (blue). (c) Circular dichroism (CD) data of the WT HMGB1 IDR peptide in the absence (black) and in the presence (gray) of 2.5 % trifluoroethanol (TFE). On the upper panel, the CD spectra are shown as the mean residue ellipticity (MRE) as a function of wavelength. On the lower panel, the high-tension voltage (HT) values are shown as a function of wavelength. Vertical dotted lines indicate the wavelength value corresponding to HT = 600 V. (d) Circular dichroism (CD) data of the Mutant HMGB1 IDR peptide in the absence (red) and in the presence (orange) of 2.5 % trifluoroethanol TFE. On the upper panel, the CD spectra are shown as the mean residue ellipticity (MRE) as a function of wavelength. On the lower panel, the high-tension voltage (HT) values are shown as a function of wavelength. Vertical dotted lines indicate the wavelength value corresponding to HT = 600 V. (e) Representative CD spectra of α-helix, β-strand and disordered proteins, shown as the mean residue ellipticity (MRE) as a function of wavelength. The data was obtained from the Protein Circular Dichroism Data Bank (PCDDB) (see Methods).