@inbook{3fbf177d6fa243219b932b2d45d4965f,
title = "Purification, crystallization and drug screening of the IRAK pseudokinases",
abstract = "Pseudokinases are emerging as critical components of cell signaling pathways. Consequently, the ability to obtain large quantities of pure protein for structural characterization and drug discovery efforts has become essential for the study of these proteins. Small molecules binding to pseudokinases may induce allosteric changes and serve as valuable tools to study the physiological roles of these {"}dead{"} enzymes. The IRAK family of kinases are key components of the innate immune response and the active IRAK family members, IRAK-1 and -4, have been extensively studied. However, the other two IRAKs, IRAK-2 and IRAK-3, are classified as pseudokinases and their detailed functions and roles remain to be described. In this chapter, we present comprehensive protocols for the purification of IRAKs, the crystallization of the pseudokinase domain of IRAK3, and a high-throughput drug screening pipeline using thermal shift and biolayer-interferometry assays to identify small molecule binders.",
keywords = "Crystallization, Drug Evaluation, Preclinical, Immunity, Innate, Signal Transduction, Protein crystallization, Protein purification, High-throughput screening, Innate immunity, IRAK3, Cell signaling, Biolayer-interferometry (BLI), Interleukin-1 receptor (IL1R), IRAK2, Toll-like receptor (TLR), Pseudokinases, Drug discovery, Thermal shift assay",
author = "Lange, {Sven M.} and Yogesh Kulathu",
note = "Funding Information: This work has been supported by Medical Research Scotland and Janssen Pharmaceutica in the form of a PhD studentship awarded to SML (PhD-770-2014). YK is supported by MRC grant MC_UU_00018/3, the EMBO Young Investigator Program and the Lister Institute of Preventive Medicine. Copyright {\textcopyright} 2022 Elsevier Inc. All rights reserved.",
year = "2022",
month = may,
day = "4",
doi = "10.1016/bs.mie.2022.03.026",
language = "English",
isbn = "9780323915410",
volume = "667",
series = "Methods in Enzymology",
publisher = "Academic Press",
pages = "101--121",
editor = "Natalia Jura and Murphy, {James M.}",
booktitle = "Pseudokinases",
}