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Peroxide-dependent sulfenylation of the EGFR catalytic site enhances kinase activity

Nature Chemical Biology volume 8, pages 57–64 (2012)Cite this article

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Abstract

Protein sulfenylation is a post-translational modification of emerging importance in higher eukaryotes. However, investigation of its diverse roles remains challenging, particularly within a native cellular environment. Herein we report the development and application of DYn-2, a new chemoselective probe for detecting sulfenylated proteins in human cells. These studies show that epidermal growth factor receptor–mediated signaling results in H2O2 production and oxidation of downstream proteins. In addition, we demonstrate that DYn-2 has the ability to detect differences in sulfenylation rates within the cell, which are associated with differences in target protein localization. We also show that the direct modification of epidermal growth factor receptor by H2O2 at a critical active site cysteine (Cys797) enhances its tyrosine kinase activity. Collectively, our findings reveal sulfenylation as a global signaling mechanism that is akin to phosphorylation and has regulatory implications for other receptor tyrosine kinases and irreversible inhibitors that target oxidant-sensitive cysteines in proteins.

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Figure 1: Cellular redox status affects EGF-mediated signaling.
Figure 2: Development and validation of probes for detecting sulfenic acid.
Figure 3: EGF-mediated ROS production and protein sulfenylation.
Figure 4: Differential sulfenylation of PTPs in EGF-treated cells.
Figure 5: EGF-mediated sulfenylation of EGFR Cys797 in cells.
Figure 6: Model for redox regulation of EGFR signaling.

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Acknowledgements

The authors acknowledge funding from the Camille Henry Dreyfus Teacher Scholar Award (to K.S.C.) and the American Heart Association Scientist Development Award (0835419N to K.S.C.). The authors also wish to thank R. Petter for helpful discussions.

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Authors and Affiliations

  1. Chemical Biology Graduate Program, University of Michigan, Ann Arbor, Michigan, USA

    Candice E Paulsen & Stephen E Leonard

  2. Department of Chemistry, University of Michigan, Ann Arbor, Michigan, USA

    Thu H Truong

  3. Department of Chemistry, The Scripps Research Institute, Jupiter, Florida, USA

    Francisco J Garcia, Arne Homann, Vinayak Gupta & Kate S Carroll

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  1. Candice E Paulsen
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Contributions

C.E.P., T.H.T. and A.H. performed cell culture and immunostaining experiments. F.J.G. performed mass spectrometry experiments. V.G. and S.E.L. performed synthetic experiments. K.S.C. designed experimental strategies with help from C.E.P. and V.G. K.S.C. and C.E.P. wrote the paper with input from all coauthors.

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Correspondence to Kate S Carroll.

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Paulsen, C., Truong, T., Garcia, F. et al. Peroxide-dependent sulfenylation of the EGFR catalytic site enhances kinase activity. Nat Chem Biol 8, 57–64 (2012). https://doi.org/10.1038/nchembio.736

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