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A yeast-based screen reveals that sulfasalazine inhibits tetrahydrobiopterin biosynthesis

Nature Chemical Biology volume 7, pages 375–383 (2011)Cite this article

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Abstract

We introduce an approach for detection of drug-protein interactions that combines a new yeast three-hybrid screening for identification of interactions with affinity chromatography for their unambiguous validation. We applied the methodology to the profiling of clinically approved drugs, resulting in the identification of previously known and unknown drug-protein interactions. In particular, we were able to identify off-targets for erlotinib and atorvastatin, as well as an enzyme target for the anti-inflammatory drug sulfasalazine. We demonstrate that sulfasalazine and its metabolites, sulfapyridine and mesalamine, are inhibitors of the enzyme catalyzing the final step in the biosynthesis of the cofactor tetrahydrobiopterin. The interference with tetrahydrobiopterin metabolism provides an explanation for some of the beneficial and deleterious properties of sulfasalazine and furthermore suggests new and improved therapies for the drug. This work thus establishes a powerful approach for drug profiling and provides new insights in the mechanism of action of clinically approved drugs.

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Figure 1: A Y3H system based on SNAP-tag labeling.
Figure 2: Illustration of the yeast-based platform for drug profiling.
Figure 3: Analysis of hits obtained from purvalanol B and erlotinib profiling.
Figure 4: Analysis of hits obtained from atorvastatin profiling.
Figure 5: Analysis of the interaction of sulfasalazine with SPR.

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Acknowledgements

We thank D. Auerbach (Dualsystems Biotech) for sharing plasmid pU6H3VSV, strain NMY51, BG-purvalanol B and cDNA libraries; N. Johnsson (Ulm University) for sharing plasmid pSH47; and O. Hantschel and G. Superti-Furga (Center for Molecular Medicine of the Austrian Academy of Sciences) for sharing plasmid pETM30-NQO2. We thank S. Moser, L. Reymond and C. Fellay for technical assistance. We acknowledge R. Chidley, M.J. Hinner and G. Lukinavicius for critically reading the manuscript. This work was supported by the Swiss National Science Foundation and the Federal Office for Professional Education and Technology.

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  1. Institute of Chemical Sciences and Engineering, Institute of Bioengineering, National Centre of Competence in Research Chemical Biology, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland

    Christopher Chidley, Hirohito Haruki, Miriam Grønlund Pedersen, Evelyne Muller & Kai Johnsson

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  1. Christopher Chidley
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Contributions

C.C., H.H. and K.J. designed experiments and analyzed data, C.C. developed and performed yeast-based screens, H.H. and C.C. performed hit validation experiments, M.G.P. measured intracellular biopterin levels, E.M. synthesized drug derivatives, and C.C. and K.J. wrote the paper.

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Correspondence to Kai Johnsson.

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The authors declare no competing financial interests.

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Supplementary Methods, Supplementary Figures 1–15 and Supplementary Tables 1 & 2 (PDF 7752 kb)

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Chidley, C., Haruki, H., Pedersen, M. et al. A yeast-based screen reveals that sulfasalazine inhibits tetrahydrobiopterin biosynthesis. Nat Chem Biol 7, 375–383 (2011). https://doi.org/10.1038/nchembio.557

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