Methods for the recombinant expression of active tyrosine kinase domains: Guidelines and pitfalls

M. Escarlet Díaz Galicia, Abdullah Aldehaiman, Seungbeom Hong, Stefan T. Arold, Raik Grunberg

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Protein tyrosine kinases (PTKs) are key signaling molecules and important drug targets. Although the efficient recombinant production of active PTKs is important for both pharmaceutical industry and academic research, most PTKs are still obtained from conventional, expensive and time-consuming insect-cell based expression. Host toxicity, kinase inactivity, insolubility and heterogeneity are among the reasons thought to preclude PTK expression in Escherichia coli. Herein we review these presumed roadblocks and their possible solutions for bacterial expression of PTKs, and give an overview on kinase activity assays. Finally, we report our experiences and observations with the kinases Src, Lyn and FAK as examples to illustrate implementation, effects and pitfalls of E. coli expression and in vitro assaying of PTKs.
Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherElsevier BV
Pages131-152
Number of pages22
ISBN (Print)9780128181171
DOIs
StatePublished - Mar 26 2019

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