Fig. 7: Location of INSR variants with impaired insulin binding amenable to monoclonal antibody activation.

A, B Manhattan plots displaying differential signalling scores for (A) mAb 83-7 and (B) mAb 83-14 compared to insulin, for all variants whose mutation severely impairs insulin signalling. Only loss-of-signalling variants with antibody signalling results from all three replicates and which scored in all five insulin signalling replicates are plotted. Black dots denote variants activatable only by the single mAb plotted. The horizontal line represents a selective sensitivity score of +0.204. D734A, a variant previously extensively studied, is labelled. Green or blue stars on X axes indicate site 1 and site 2 residues respectively. Areas identified on variant effect maps as showing selective activation by antibodies are boxed for reference, and a frequency histogram indicating counts of potential target variants in a sliding 10 residue window is superimposed. C 4 insulin-bound INSR structure (PDB: 6PXV), coloured by mean sensitivity scores for mAb 83-7 and mAb 83-14. Insulin molecules are green. The left hand box shows a zoomed in view of insulin bound to binding site 1, and the right hand box shows the view of the structure from above. D Scatter plot comparing selective sensitivity score for mAbs 83-7 and 83-14, divided into quadrants according to the arbitrary threshold score +0.204 used in (A, B). Selected variants are labelled, and proportions of variants in each quadrant are indicated.