Identifying physical interactions between proteins and other molecules is a critical aspect of biological analysis. Here we describe PLATO, an in vitro method for mapping such interactions by affinity enrichment of a library of full-length open reading frames displayed on ribosomes, followed by massively parallel analysis using DNA sequencing. We demonstrate the broad utility of the method for human proteins by identifying known and previously unidentified interacting partners of LYN kinase, patient autoantibodies, and the small-molecules gefitinib and dasatinib.
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We would like to thank K. Waraska, M. Cicero, S. Alian and A. Gagne for assistance with Illumina sequencing, and J. Laserson for statistical advice. Thanks to D. Zhu for help with synthesis of biotin-dasatinib, which was partially supported by US National Institutes of Health U54 CA156734 to the University of Massachusetts Boston–Dana-Farber Harvard Cancer Center U54 Comprehensive Partnership (Project 3, Co-PIs: N.S. Gray, W. Zhang, and P.L. Yang). We also thank N. Gray at Harvard Medical School for valuable advice, and H. Varmus at the National Cancer Institute for providing gefitinib reagents and advice. This work was supported in part by NIH grant 3P30CA023100-25S8 to S.K. S.J.E. is an investigator with the Howard Hughes Medical Institute.
The authors declare no competing financial interests.
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Zhu, J., Larman, H., Gao, G. et al. Protein interaction discovery using parallel analysis of translated ORFs (PLATO). Nat Biotechnol 31, 331–334 (2013). https://doi.org/10.1038/nbt.2539
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