Deep quantification of substrate turnover defines protease subsite cooperativity

ABSTRACT: Substrate specificity determines protease functions in physiology, and in clinical and biotechnological application, yet quantitative cleavage information is often unavailable, biased, or limited to a small number of events. Here, we develop qPISA (quantitative Protease specificity Inference from Substrate Analysis) to study Dipeptidyl Peptidase Four (DPP4), a key regulator of blood glucose levels. We use mass spectrometry to quantify >40,000 peptides from a complex, commercially available peptide mixture. By analyzing changes in substrate levels quantitatively instead of focusing on qualitative product identification through a binary classifier, we can reveal cooperative interactions within DPP4's active pocket and derive a sequence motif that predicts activity quantitatively. qPISA di

SUBMITTER: Rajani, Kanth Gudipati 

PROVIDER: S-SCDT-10_1038-S44320-024-00071-4 | biostudies-other |

REPOSITORIES: biostudies-other