Titlebook: Deubiquitinases; Methods and Protocol Julie Maupin-Furlow,Mariola J. Edelmann Book 2023 The Editor(s) (if applicable) and The Author(s), un

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https://doi.org/10.1007/978-3-030-21406-7e describe a step-by-step experimental procedure to assess the activity of PLpro in vitro using a suite of activity-based probes (ABPs) and fluorescent substrates and how they can be applied as fast and yet sensitive methods to calculate kinetic parameters.

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Julie Maupin-Furlow,Mariola J. EdelmannIncludes cutting-edge methods and protocols.Provides step-by-step detail essential for reproducible results.Contains key notes and implementation advice from the experts

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Activity-Based Protein Profiling (ABPP) for Cellular Deubiquitinase (DUB) and Inhibitor Profiling a probes, with immunoblotting and mass spectrometry proteomics-based readouts. Different types of activity-based protein profiling (ABPP) for studying the potency and selectivity of DUB inhibitors are outlined here, including the standard ABPP, the deep DUBome ABPP, and the ABPP-HT (high-throughput compatible).

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https://doi.org/10.1007/978-1-0716-2803-4DUB inhibitors; Next-generation phage display; poly-Ub chain; therapeutic opportunities v; innate immuni

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High Mountain Hazards in Uttarakhandd catalytic triad rearrangement, where ubiquitin-binding helps the DUB adopt an active conformation for catalysis. The crystal structure of the apo form of such a DUB, when not bound to ubiquitin, reveals an inactive conformation of the catalytic residues, necessitating the structure of the ubiquiti

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Navdeep Agrawal,Jagabandhu Dixitprecise characterization of both enzyme and DUB inhibitor. These probes are compatible with most plate readers allowing for rapid, facile fluorometric analysis of DUB activity. DUB activity can be measured in purified enzyme reactions, in cell lysates, or in intact cells depending upon the choice of