Fig 2. In silico saturation mutagenesis analysis of ROMK variants.

The predicted pathogenicity probabilities obtained with Rhapsody are shown in a heat map (divided in two parts) with a color code ranging from red (deleterious, >0.5) to blue (neutral, ≤0.5). Light yellow entries correspond to wild-type (WT) amino acids. The bottom panels corresponding to each heat map represent the residue-averaged pathogenicities predicted by Rhapsody (red), EVmutation (green) and PolyPhen-2 (blue). The pink shade indicates the range of Rhapsody results for the 19 specific substitutions for each residue. The locations of 31 variants selected for experimental testing and two controls are marked by black squares on the map, and callouts colored by the measured phenotype, with the same color scheme as in Figs 3 and 5. On the right, the residue-averaged predictions from Rhapsody are displayed for a single monomer in the human ROMK1 homology model using the same color code as in the map.