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Comprehensive assay of kinase catalytic activity reveals features of kinase inhibitor selectivity

Nature Biotechnology volume 29, pages 1039–1045 (2011)Cite this article

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Abstract

Small-molecule protein kinase inhibitors are widely used to elucidate cellular signaling pathways and are promising therapeutic agents. Owing to evolutionary conservation of the ATP-binding site, most kinase inhibitors that target this site promiscuously inhibit multiple kinases. Interpretation of experiments that use these compounds is confounded by a lack of data on the comprehensive kinase selectivity of most inhibitors. Here we used functional assays to profile the activity of 178 commercially available kinase inhibitors against a panel of 300 recombinant protein kinases. Quantitative analysis revealed complex and often unexpected interactions between protein kinases and kinase inhibitors, with a wide spectrum of promiscuity. Many off-target interactions occur with seemingly unrelated kinases, revealing how large-scale profiling can identify multitargeted inhibitors of specific, diverse kinases. The results have implications for drug development and provide a resource for selecting compounds to elucidate kinase function and for interpreting the results of experiments involving kinase inhibitors.

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Figure 1: Large-scale kinase-inhibitor interaction analysis.
Figure 2: Comparison of functional inhibition data generated in this study with previous kinase-inhibitor interaction profiling studies.
Figure 3: Kinase selectivity.
Figure 4: Kinase inhibitor selectivity.
Figure 5: Uni-specific kinase inhibitors.

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Acknowledgements

We gratefully acknowledge B. Turk, A. Andrews and members of the Peterson laboratory for comments on the manuscript and R. Hartman of Reaction Biology Corp. for developing the Kinase Inhibitor Resource (KIR) web application tool. This work was supported by a W.W. Smith Foundation Award, funding from the Keystone Program in Head and Neck Cancer of Fox Chase Cancer Center and by US National Institutes of Health awards RO1 GM083025 to J.R.P. and P30 CA006927 to Fox Chase Cancer Center. HotSpot technology development was partially supported by the US National Institutes of Health (RO1 HG003818 and R44 CA114995 to H.M.).

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Authors and Affiliations

  1. Cancer Biology Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA

    Theonie Anastassiadis, Karthik Devarajan & Jeffrey R Peterson

  2. Reaction Biology Corporation, Malvern, Pennsylvania, USA

    Sean W Deacon & Haiching Ma

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  1. Theonie Anastassiadis
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  2. Sean W Deacon
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  5. Jeffrey R Peterson
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Contributions

The study was conceived by J.R.P., S.W.D. and H.M., experimental data was collected by S.W.D., statistical analysis was performed by K.D., data were analyzed by T.A. and J.R.P. with input from S.W.D. and H.M., and the manuscript was written by J.R.P. with input from the other authors.

Corresponding author

Correspondence to Jeffrey R Peterson.

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Competing interests

S.W.D. and H.M. are current employees of Reaction Biology Corporation.

Supplementary information

Supplementary Text and Figures

Supplementary Tables 1,2,4,5 and Supplementary Figures 1–5 (PDF 10400 kb)

Supplementary Table 3

Complete pairwise kinase-compound activity dataset. (XLS 967 kb)

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Anastassiadis, T., Deacon, S., Devarajan, K. et al. Comprehensive assay of kinase catalytic activity reveals features of kinase inhibitor selectivity. Nat Biotechnol 29, 1039–1045 (2011). https://doi.org/10.1038/nbt.2017

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