ID
Ilya V. Demidyuk
  • Faculty, Institute of Molecular Genetics of National Research Centre “Kurchatov Institute”, Moscow
Research fields
  • Molecular Biology
Assay for Protealysin-like Protease Inhibitor Activity
Authors:  Igor M. Berdyshev, Maria A. Karaseva and Ilya V. Demidyuk, date: 10/05/2022, view: 840, Q&A: 0

Here, we present the first quantitative method for the activity analysis of protealysin-like protease (PLP) inhibitors. This approach is based on a previously developed method for protealysin activity determination by hydrolysis of internally quenched fluorescent peptide substrate 2-aminobenzoyl-L-arginyl-L-seryl-L-valyl-L-isoleucyl-L-(ϵ-2,4-dinitrophenyl)lysine. In this protocol, we significantly reduced enzyme concentration and introduced some minor modifications to decrease variation between replicates. The protocol was validated using emfourin, a novel proteinaceous metalloprotease inhibitor. Data obtained demonstrates that the developed assay method is an affordable approach for characterizing and screening various PLP inhibitors.


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