Mapping specificity, cleavage entropy, allosteric changes and substrates of blood proteases in a high-throughput screen.

ABSTRACT: Proteases are among the largest protein families and critical regulators of biochemical processes like apoptosis and blood coagulation. Knowledge of proteases has been expanded by the development of proteomic approaches, however, technology for multiplexed screening of proteases within native environments is currently lacking behind. Here we introduce a simple method to profile protease activity based on isolation of protease products from native lysates using a 96FASP filter, their analysis in a mass spectrometer and a custom data analysis pipeline. The method is significantly faster, cheaper, technically less demanding, easy to multiplex and produces accurate protease fingerprints. Using the blood cascade proteases as a case study, we obtain protease substrate profiles that can be used t

SUBMITTER: Uliana F 

PROVIDER: S-EPMC7966775 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature