Abstract
The chemical modification of proteins is an enabling technology for many scientific fields, including chemical biology, biophysics, bioengineering and materials science. These methods allow the attachment of strategically selected detection probes, polymers, drug molecules and analysis platforms. However, organic reactions that can proceed under conditions mild enough to maintain biomolecular function are limited. Even more rare are chemical strategies that can target a single site, leading to products with uniform properties and optimal function. We present a versatile method for the selective modification of protein N termini that does not require any genetic engineering of the protein target. This reaction is demonstrated for 12 different proteins, including the soluble domain of the human estrogen receptor. The function of this protein was confirmed through the binding of a fluorescent estrogen mimic, and the modified protein was explored as a prototype for the detection of endocrine-disrupting chemicals in water.
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Acknowledgements
The development of this reaction was supported by the Energy Biosciences Institute at the University of California–Berkeley. The application of the reaction to the human estrogen receptor was supported by the US National Science Foundation (CHE-1059083 and CHE-1413666). J.I.M. was supported by the Berkeley Chemical Biology Graduate Program (National Research Service Award Training grant 1 T32 GMO66698). H.K.M. was supported by the Villum Kann Rasmussens Foundation as well as the Laboratory Directed Research and Development Program at Lawrence Berkeley National Labs. We would like to acknowledge M. Dedeo for the tobacco mosaic virus coat protein, L. Witus for the GFP and A. Obermeyer for X-ADSWAG peptides (all at the University of California-Berkeley).
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J.I.M. and M.B.F. conceived the project. J.I.M. developed the bioconjugation reaction and synthesized the 2PCA derivatives. H.K.M. performed the protein immobilization and binding experiments. H.K.M. and T.M. performed the expression and purification of uteroglobin and the human estrogen receptor. J.I.M. and M.B.F. wrote the manuscript. All of the authors reviewed and contributed to the manuscript.
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Supplementary Results, Supplementary Figures 1–13 and Supplementary Note. (PDF 1934 kb)
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MacDonald, J., Munch, H., Moore, T. et al. One-step site-specific modification of native proteins with 2-pyridinecarboxyaldehydes. Nat Chem Biol 11, 326–331 (2015). https://doi.org/10.1038/nchembio.1792
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DOI: https://doi.org/10.1038/nchembio.1792
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