A structure-based computational mutagenesis elucidates the spectrum of stability-activity relationships in proteins

Protein engineering experiments involving single amino acid substitutions are routinely implemented for the analysis of protein structure, stability, and function. The resulting change in just one of these characteristics relative to the native protein constitutes the focus of any single study, as is the case with predictive computational models developed for the same purpose. Other than investigations into stability-activity trade-offs specifically resulting from active site residue replacements in a few enzymes, a literature survey fails to reveal a comprehensive analysis of stability-activity relationships in proteins upon mutation. Here, we employ a computational mutagenesis for quantifying overall protein structural change upon mutation, which is applied to a dataset of 938 single residue replacements distributed at positions throughout twenty diverse proteins. These mutants are selected based on the availability of both experimental stability and activity change data, and their structural change data are used to characterize the full range of stability-activity relationships.