Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Brief Communication
  • Published:

A new screening assay for allosteric inhibitors of cSrc


Targeting kinases outside the highly conserved ATP pocket is thought to be a promising strategy for overcoming bottlenecks in kinase inhibitor research, such as limited selectivity and drug resistance. Here we report the development and application of a direct binding assay to detect small molecules that stabilize the inactive conformation of the tyrosine kinase cSrc. Protein X-ray crystallography validated the assay results and confirmed an exclusively allosteric binding mode.

This is a preview of subscription content, access via your institution

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Schematic diagram of the fluorescent-labeled kinase assay for detecting allosteric inhibitors.
Figure 2: Screening of a compound library reveals pyrazoloureas binding to cSrc and p38α.
Figure 3: Crystal structures of cSrc in complex with type III inhibitors.

Similar content being viewed by others

Accession codes


Protein Data Bank


  1. Gschwind, A., Fischer, O.M. & Ullrich, A. Nat. Rev. Cancer 4, 361–370 (2004).

    Article  CAS  Google Scholar 

  2. Zhang, J., Yang, P.L. & Gray, N.S. Nat. Rev. Cancer 9, 28–39 (2009).

    Article  Google Scholar 

  3. Backes, A.C., Zech, B., Felber, B., Klebl, B. & Müller, G. Expert Opin. Drug Discovery 3, 1409–1425 (2008).

    Article  CAS  Google Scholar 

  4. Backes, A.C., Zech, B., Felber, B., Klebl, B. & Müller, G. Expert Opin. Drug Discovery 3, 1427–1449 (2008).

    Article  CAS  Google Scholar 

  5. Liu, Y. & Gray, N.S. Nat. Chem. Biol. 2, 358–364 (2006).

    Article  CAS  Google Scholar 

  6. Adrián, F.J. et al. Nat. Chem. Biol. 2, 95–102 (2006).

    Article  Google Scholar 

  7. Regan, J. et al. J. Med. Chem. 45, 2994–3008 (2002).

    Article  CAS  Google Scholar 

  8. Copeland, R.A., Pompliano, D.L. & Meek, T.D. Nat. Rev. Drug Discov. 5, 730–739 (2006).

    Article  CAS  Google Scholar 

  9. Wood, E.R. et al. Cancer Res. 64, 6652–6659 (2004).

    Article  CAS  Google Scholar 

  10. Daub, H., Specht, K. & Ullrich, A. Nat. Rev. Drug Discov. 3, 1001–1010 (2004).

    Article  CAS  Google Scholar 

  11. Annis, D.A., Nazef, N., Chuang, C.C., Scott, M.P. & Nash, H.M. J. Am. Chem. Soc. 126, 15495–15503 (2004).

    Article  CAS  Google Scholar 

  12. Jacobs, M.D., Caron, P.R. & Hare, B.J. Proteins 70, 1451–1460 (2008).

    Article  CAS  Google Scholar 

  13. Pargellis, C. et al. Nat. Struct. Biol. 9, 268–272 (2002).

    Article  CAS  Google Scholar 

  14. Yeatman, T.J. Nat. Rev. Cancer 4, 470–480 (2004).

    Article  CAS  Google Scholar 

  15. Michalczyk, A. et al. Bioorg. Med. Chem. 16, 3482–3488 (2008).

    Article  CAS  Google Scholar 

  16. Dar, A.C., Lopez, M.S. & Shokat, K.M. Chem. Biol. 15, 1015–1022 (2008).

    Article  CAS  Google Scholar 

  17. Seeliger, M.A. et al. Structure 15, 299–311 (2007).

    Article  CAS  Google Scholar 

  18. Dumas, J. et al. Bioorg. Med. Chem. Lett. 10, 2051–2054 (2000).

    Article  CAS  Google Scholar 

  19. Regan, J. et al. J. Med. Chem. 46, 4676–4686 (2003).

    Article  CAS  Google Scholar 

  20. Wan, P.T. et al. Cell 116, 855–867 (2004).

    Article  CAS  Google Scholar 

  21. Levinson, N.M. et al. PLoS Biol. 4, e144 (2006).

    Article  Google Scholar 

Download references


We thank the Dortmund Protein Facility for cloning, expressing and purifying the chicken cSrc and human p38α mutant variants for labeling. We thank M. Weyand, W. Blanckenfeldt, E. Hofmann, I. Vetter and beamline scientists at the Swiss Light Source of the Paul Scherrer Institute for expert assistance during data collection. We thank P. Janning for her assistance with the HPLC and MS analysis of the fluorescent-labeled and unlabeled cSrc, and we thank H. Waldmann and K. Shokat for helpful discussions. J.R.S. was funded by the Alexander von Humboldt Foundation. This work was supported by the German Federal Ministry for Education and Research through the German National Genome Research Network-Plus (NGFN-Plus) (grant number BMBF 01GS08102). Schering Plough, Bayer-Schering Pharma, Merck-Serono and Bayer CropScience are thanked for financial support.

Author information

Authors and Affiliations



J.R.S. designed the p38α and cSrc constructs for labeling, developed the fluorescent-labeled kinase assay system and determined compound Kd, kon and koff values. S.K. overexpressed, purified and crystallized cSrc. C.G. determined the cocrystal structures. M.R. determined IC50 values. M.G. synthesized the compounds with assistance from S.K. and H.B.R. The manuscript was prepared by J.R.S. with assistance by all co-authors. D.R. designed the experiments.

Corresponding author

Correspondence to Daniel Rauh.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–4, Supplementary Tables 1 and 2, and Supplementary Methods (PDF 874 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Simard, J., Klüter, S., Grütter, C. et al. A new screening assay for allosteric inhibitors of cSrc. Nat Chem Biol 5, 394–396 (2009).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

This article is cited by


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing