@inbook{9d9d553625f840a5a14270838c2895f1,
title = "High-Throughput MALDI-TOF Mass Spectrometry-Based Deubiquitylating Enzyme Assay for Drug Discovery",
abstract = "Ubiquitylation (or ubiquitination) is the reversible conjugation of a 76-amino-acid polypeptide (ubiquitin) to a target protein to modulate various biological processes. Deubiquitylating enzymes (DUBs) are a class of enzymes that specifically remove ubiquitin from a substrate. In recent years DUBs have garnered significant attention as a new class of targets in multiple therapeutic areas. The recent development of high-throughput Matrix-Assisted Laser Desorption/Ionization-Time of Flight mass spectrometry (MALDI-TOF MS) has provided new tools to perform drug discovery screening. Here we present a facile and high-throughput step-by-step protocol of the MALDI-TOF MS-based DUB assay for screening the activity of DUBs in vitro. In a MALDI-TOF DUB assay, we quantitate the amount of mono-ubiquitin generated by the in vitro cleavage of ubiquitin chains. The presented protocol takes advantage of nanoliter dispensing robotics and automated MALDI-TOF MS analysis to screen large and diverse compound libraries.",
keywords = "Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods, High-Throughput Screening Assays/methods, Drug Discovery/methods, Ubiquitin/metabolism, Enzyme Assays, Ubiquitin, DUBs, Deubiquitylating enzyme, High-throughput screening, MALDI-TOF mass spectrometry, Drug discovery",
author = "{De Cesare}, Virginia and Paul Davies",
note = "Copyright: {\textcopyright} 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2023",
doi = "10.1007/978-1-0716-2803-4_8",
language = "English",
isbn = "9781071628027",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "123--134",
editor = "Julie Maupin-Furlow and Edelmann, {Mariola J.}",
booktitle = "Deubiquitinases",
address = "United States",
edition = "1",
}