Feb 3, 2016

An open library of human kinase domain constructs for automated bacterial expression

BioRxiv : the Preprint Server for Biology
Daniel L PartonJohn D. Chodera


Kinases play a critical role in many cellular signaling pathways and are dysregulated in a number of diseases, such as cancer, diabetes, and neurodegeneration. Since the FDA approval of imatinib in 2001, therapeutics targeting kinases now account for roughly 50% of current cancer drug discovery efforts. The ability to explore human kinase biochemistry, biophysics, and structural biology in the laboratory is essential to making rapid progress in understanding kinase regulation, designing selective inhibitors, and studying the emergence of drug resistance. While insect and mammalian expression systems are frequently used for the expression of human kinases, bacterial expression systems are superior in terms of simplicity and cost-effectiveness but have historically struggled with human kinase expression. Following the discovery that phosphatase coexpression could produce high yields of Src and Abl kinase domains in bacterial expression systems, we have generated a library of 52 His-tagged human kinase domain constructs that express above 2 μg/mL culture in a simple automated bacterial expression system utilizing phosphatase coexpression (YopH for Tyr kinases, Lambda for Ser/Thr kinases). Here, we report a structural bioinformatic...Continue Reading

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Mentioned in this Paper

United States Food and Drug Administration
Biochemical Pathway
Antineoplastic Agents
Lambda (Sutural Junction)
Phosphoric Monoester Hydrolases
High Throughput Screening
YopH protein, Yersinia
Nerve Degeneration
High Throughput Technology

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