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Proteome-wide drug and metabolite interaction mapping by thermal-stability profiling

Nature Methods volume 12, pages 10551057 (2015) | Download Citation

Abstract

Thermal stabilization of proteins after ligand binding provides an efficient means to assess the binding of small molecules to proteins. We show here that in combination with quantitative mass spectrometry, the approach allows for the systematic survey of protein engagement by cellular metabolites and drugs. We profiled the targets of the drugs methotrexate and (S)-crizotinib and the metabolite 2′3′-cGAMP in intact cells and identified the 2′3′-cGAMP cognate transmembrane receptor STING, involved in immune signaling.

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Acknowledgements

We are grateful to W. Berger (Institute of Cancer Research, Vienna, Austria) for providing SW480 cells and P. Majek (CeMM, Vienna, Austria) for assistance with data processing. This work was supported by the Austrian Academy of Sciences, the European Union (FP7 259348, ASSET) and the Austrian Science Fund (FWF F4711, MPN).

Author information

Author notes

    • Kilian V M Huber
    • , Chris Soon Heng Tan
    •  & Jacques Colinge

    Present addresses: Structural Genomics Consortium, University of Oxford, Oxford, UK and Target Discovery Institute, University of Oxford, Oxford, UK (K.V.M.H.); Institute of Molecular & Cell Biology, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore and Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), Singapore, Singapore (C.S.H.T.); Institut de Recherche en Cancérologie de Montpellier, Inserm U1194, Université Montpellier, Montpellier, France (J.C.).

Affiliations

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

    • Kilian V M Huber
    • , Karin M Olek
    • , André C Müller
    • , Chris Soon Heng Tan
    • , Keiryn L Bennett
    • , Jacques Colinge
    •  & Giulio Superti-Furga
  2. Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria.

    • Giulio Superti-Furga

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Contributions

K.V.M.H. designed experiments and jointly performed them with K.M.O.; A.C.M. and K.L.B. performed mass spectrometry; C.S.H.T. and J.C. performed bioinformatics analysis; and K.V.M.H. and G.S.-F. conceived the study and wrote the manuscript. All authors contributed to the discussion of results and participated in preparation of the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Kilian V M Huber or Jacques Colinge or Giulio Superti-Furga.

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DOI

https://doi.org/10.1038/nmeth.3590

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