Engineered Chymotrypsins and Uses Thereof
Engineering proteolytic specificity to enable detection of post-translationally modified (PTM) amino acids has been demonstrated for the bacterial proteases OmpT and subtilisin, and more recently trypsin. However, a need exists to broaden the toolkit of proteases that can be used for mass-spectrometry based proteases. This invention is an engineered form of Chymotrypsin with substrate specificity with an expanded specificity for Asparagine(Asn) containing peptides and proteins. This modified chymotrypsin proteolyzes after peptides and polypeptides C-terminal to Asn with a kcat/KM of at least 0.5 M-1s-1.
Associate Professor, Chemical Engineering