Activity-Based Protein Profiling (ABPP) of Cellular DeISGylating Enzymes and Inhibitor Screening.

Simeon D Draganov, Hannah B L Jones, Qura Shah, Benedikt M Kessler, Adán Pinto-Fernández

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Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 authors.

Simeon D DraganovChinese Academy of Medical Sciences Oxford Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK. simeon.draganov@ndm.ox.ac.uk.

Hannah B L JonesTarget Discovery Institute, Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, UK.

Qura ShahLife Sciences Solutions, Thermo Fisher Scientific, Paisley, UK.

Benedikt M KesslerChinese Academy of Medical Sciences Oxford Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK.

Adán Pinto-FernándezChinese Academy of Medical Sciences Oxford Institute, Nuffield Department of Medicine, University of Oxford, Oxford, UK. adan.pintofernandez@ndm.ox.ac.uk.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

A detailed methodology platform is described for activity-based protein profiling (ABPP) of cellular deISGylating enzymes using a specific activity-based interferon-stimulated gene 15 (ISG15) probe. Manual and semi-automated workflows for medium- to high-throughput applications are outlined in this chapter, with western blotting and proteomics-based techniques as the main readouts. This methodology informs us of endogenous deISGylating enzyme expression and activity in a cellular context, including USP18, the type I interferon (IFN-I)-inducible deISGylase, and several constitutively expressed deubiquitinases (DUBs), such as USP5, USP14, USP16, and USP36, that exert cross-reactivity to ISG15. ISG15-ABPP also enables the identification and characterization of potent and selective deISGylating enzyme modulators.

Indexed as

Deubiquitinating EnzymesEnzyme InhibitorsProteomicsBlotting, WesternCytokinesHigh-Throughput Screening AssaysHumansUbiquitinsCytokinesDeubiquitinating EnzymesEnzyme InhibitorsISG15 protein, humanUbiquitinsActivity-based probesActivity proteomicsChemical biologyDeISGylating enzymesDeISGylationDrug discoveryISG15PharmacologyProteomics

Identifiers

PMID40515982

DOI10.1038/s41592-019-0638-x

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.