Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Protein interaction discovery using parallel analysis of translated ORFs (PLATO)


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.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Parallel analysis of translated ORFs (PLATO).
Figure 2: Identification of known and previously undescribed interactions using PLATO.


  1. Faix, P.H. et al. Biotechniques 36, 1018–1022, 1024, 1026–1029 (2004).

    CAS  Article  Google Scholar 

  2. Zhu, H., Bilgin, M. & Snyder, M. Annu. Rev. Biochem. 72, 783–812 (2003).

    CAS  Article  Google Scholar 

  3. Jeong, J.S. et al. Mol. Cell Proteomics 11, O111.016253 (2012).

    Article  Google Scholar 

  4. Lamesch, P. et al. Genomics 89, 307–315 (2007).

    CAS  Article  Google Scholar 

  5. Amstutz, P., Binz, H.K., Zahnd, C. & Plückthun,, A. Ribosome display: in vitro selection of protein-protein interactions in Cell Biology—A Laboratory Handbook (ed. Celis, J.) Vol. 1, 3rd Ed., 497–509 (Elsevier Academic Press, 2006).

  6. Boggon, T.J. & Eck, M.J. Oncogene 23, 7918–7927 (2004).

    CAS  Article  Google Scholar 

  7. Weng, Z. et al. Mol. Cell Biol. 14, 4509–4521 (1994).

    CAS  Article  Google Scholar 

  8. Samuels, A.L., Klinken, S.P. & Ingley, E. Blood 113, 3845–3856 (2009).

    CAS  Article  Google Scholar 

  9. Larman, H.B. et al. Nat. Biotechnol. 29, 535–541 (2011).

    CAS  Article  Google Scholar 

  10. Carthagena, L. et al. PLoS ONE 4, e4894 (2009).

    Article  Google Scholar 

  11. Brehmer, D. et al. Cancer Res. 65, 379–382 (2005).

    CAS  PubMed  Google Scholar 

Download references


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.

Author information

Authors and Affiliations



S.J.E. and H.B.L. conceived and supervised the project. pRD human ORFeome library was constructed by J.Z., and characterized by J.Z. and H.B.L. The PLATO protocol was developed by H.B.L. and J.Z. Clinical evaluations and patient sample acquisitions were performed by S.K. Statistical analysis was performed by U.L. under the supervision of G.C. R.S. provided gefitinib-conjugated beads. PLATO candidates were confirmed by J.Z. and G.G. A.C. provided support for the validation of LYN binding candidates. N.P. provided support for the validation of PND autoantigen candidates. Z.Z. and W.Z. provided biotin-dasatinib. The manuscript was prepared by H.B.L. and J.Z., and edited by S.J.E.

Corresponding author

Correspondence to Stephen J Elledge.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–6 and Supplementary Methods (PDF 3363 kb)

Supplementary Tables 1-4

ORFeome enrichment by GST-LYN, ORFeome enrichment by PND patient autoantibodies, ORFeome enrichment by dasatinib, Sequences of DNA primers used in this study (XLSX 101 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Zhu, J., Larman, H., Gao, G. et al. Protein interaction discovery using parallel analysis of translated ORFs (PLATO). Nat Biotechnol 31, 331–334 (2013).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

Further reading


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing