Prediction of Function Determining and Buried Residues Through Analysis of Saturation Mutagenesis Datasets

Mutational scanning can be used to probe effects of large numbers point mutations on protein function. Positions affected by mutation are primarily at either buried or exposed residues directly involved in function, hereafter designated as active-site residues. In the absence prior structural information, it has not been easy distinguish between these two categories We curated and analyzed a set twelve published deep mutational datasets. The analysis revealed differential patterns sensitivity substitution preferences positions. Prediction buried-sites solely from data was facilitated incorporating predicted sequence-based accessibility values. For we observed mean sensitivity, specificity accuracy 61, 90 88% respectively. corresponding figures were 59, 84% while for non 98, 44 82% also identified positions which did follow general trends might require further experimental re-validation. This highlights ability scans provide important functional insights, even three-dimensional structures determined using conventional structure determination techniques, discuss some limitations methodology.