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Light-assisted small-molecule screening against protein kinases

Nature Chemical Biology volume 11, pages 952–954 (2015)Cite this article

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Abstract

High-throughput live-cell screens are intricate elements of systems biology studies and drug discovery pipelines. Here, we demonstrate an optogenetics-assisted method that avoids the need for chemical activators and reporters, reduces the number of operational steps and increases information content in a cell-based small-molecule screen against human protein kinases, including an orphan receptor tyrosine kinase. This blueprint for all-optical screening can be adapted to many drug targets and cellular processes.

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Figure 1: All-optical screen against RTKs and the MAPK/ERK pathway.
Figure 2: Optogenetics-enabled, internally referenced measurement of the MAPK/ERK pathway.

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Acknowledgements

We thank M. Ernst (Medical University of Vienna) for fruitful discussions, M. Spanova (IST Austria) for technical assistance, R. Chait (IST Austria) for photography, X. Amouretti and B. Harris (BioTek) for hardware specifications, R. Stahel (University of Zurich) for mesothelioma cells, and D.M. Chudakov (Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry)/Evrogen (Moscow) for mKate2. This work was supported by grants from the European Union Seventh Framework Programme (CIG-303564 to H.J. and ERC-StG-311166 to S.M.B.N.), the Human Frontier Science Program (RGY0084_2012 to H.J.) and the Herzfelder Foundation (to M.G.). A.I.-P. was supported by a Ramon Areces fellowship, and E.R. by the graduate program MolecularDrugTargets (Austrian Science Fund (FWF): W 1232) and a FemTech fellowship (3580812 Austrian Research Promotion Agency).

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Author notes

  1. Sebastian M B Nijman

    Present address: Present address: Ludwig Institute for Cancer Research, Headington, Oxford, UK.,

Authors and Affiliations

  1. Institute of Science and Technology Austria (IST Austria), Klosterneuburg, Austria

    Álvaro Inglés-Prieto, Eva Reichhart, Matthias Nowak & Harald Janovjak

  2. CeMM–Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria

    Markus K Muellner & Sebastian M B Nijman

  3. Department of Medicine I, Institute of Cancer Research, Comprehensive Cancer Center Vienna, Medical University of Vienna, Vienna, Austria

    Michael Grusch

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  1. Álvaro Inglés-Prieto
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Contributions

A.I.-P. designed, performed and analyzed all-optical experiments. E.R. designed, performed and analyzed internal reference experiments. M.K.M., M.N., S.M.B.N. and M.G. designed inhibitor library and experiments and provided reagents. H.J. conceived and supervised the project and designed and analyzed experiments. A.I.-P., E.R. and H.J. wrote the paper.

Corresponding author

Correspondence to Harald Janovjak.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Figures 1–12 and Supplementary Tables 1–3. (PDF 1461 kb)

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Inglés-Prieto, Á., Reichhart, E., Muellner, M. et al. Light-assisted small-molecule screening against protein kinases. Nat Chem Biol 11, 952–954 (2015). https://doi.org/10.1038/nchembio.1933

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