Methods for the recombinant expression of active tyrosine kinase domains: Guidelines and pitfalls
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KAUSTRep_Diaz-Galicia_Review.pdf
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Accepted Manuscript
Type
Book ChapterAuthors
Díaz Galicia, M. EscarletAldehaiman, Abdullah

Hong, Seungbeom
Arold, Stefan T.

Grunberg, Raik
KAUST Department
Biological and Environmental Sciences and Engineering (BESE) DivisionBioscience Program
Computational Bioscience Research Center (CBRC)
Computer, Electrical and Mathematical Sciences and Engineering (CEMSE) Division
KAUST Grant Number
URF/1/2602-01FCC/1/1976-21
Date
2019-03-26Online Publication Date
2019-03-26Print Publication Date
2019Permanent link to this record
http://hdl.handle.net/10754/631891
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Show full item recordAbstract
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.Citation
Díaz Galicia ME, Aldehaiman A, Hong S, Arold ST, Grünberg R (2019) Methods for the recombinant expression of active tyrosine kinase domains: Guidelines and pitfalls. Methods in Enzymology. Available: http://dx.doi.org/10.1016/bs.mie.2019.02.027.Sponsors
This research was supported by the King Abdullah University of Science and Technology (KAUST) through the baseline fund and the Award No. URF/1/2602-01 and FCC/1/1976-21 from the Office of Sponsored Research (OSR). Supplemental data, detailed methods and complete sequences are available on http://github.com/strubelab/kinaseexpress.Publisher
Elsevier BVJournal
Methods in EnzymologyAdditional Links
https://www.sciencedirect.com/science/article/pii/S0076687919300461ae974a485f413a2113503eed53cd6c53
10.1016/bs.mie.2019.02.027