Figure 7.

AlphaFold2-ColabFold ensembles and high sequence conservation in the FF4 subdomain. (A, B) The 80 superimposed AlphaFold2-predicted structures of the WT (A), and the relative orientations of the charge troika, are depicted in sticks to highlight the two rotameric states of K113 (B). (C, D) The superimposed predicted structures of K113E mutant (C), and the orientations of the corresponding troika residues at the interface (D). (E) The χ₁ and χ₂ rotameric dihedral angle distributions for residue 113 of WT (red) and the mutant (blue) show two different orientations in the former construct. The arrows represent the orientation present in the rank 1 model, which was used for all structure-based predictions. (F) Shannon entropy as a function of sequence index, calculated from multiple sequence alignment (MSA) of FF34 orthologs. The orange circles represent the troika residues. (G) The sequence logo obtained from the MSA shows the conservation of troika residues.