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

Jian Zhu; H Benjamin Larman; Geng Gao; Romel Somwar; Zijuan Zhang; Uri Laserson; Alberto Ciccia; Natalya Pavlova; George Church; Wei Zhang; Santosh Kesari; Stephen J Elledge

doi:10.1038/nbt.2539

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

Nat Biotechnol. 2013 Apr;31(4):331-334. doi: 10.1038/nbt.2539. Epub 2013 Mar 17.

Authors

Jian Zhu^#^{1

2

3}, H Benjamin Larman^#^{1

4

5

2

3}, Geng Gao^{1

2

3}, Romel Somwar⁶, Zijuan Zhang⁷, Uri Laserson^{4

2

8}, Alberto Ciccia^{1

2

3}, Natalya Pavlova^{1

2

3}, George Church^{4

2}, Wei Zhang⁷, Santosh Kesari⁹, Stephen J Elledge^{1

2

3}

Affiliations

¹ Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Boston, MA.
² Department of Genetics, Harvard University Medical School, Boston, MA.
³ Howard Hughes Medical Institute.
⁴ Harvard-MIT Division of Health Sciences and Technology, Cambridge, MA.
⁵ Department of Materials Science and Engineering, Massachusetts Institute of Technology, Cambridge, MA.
⁶ Human Oncology and Pathogenesis Program, Memorial Sloan-Kettering Cancer Center, New York, NY.
⁷ Department of Chemistry, University of Massachusetts Boston, Boston, MA.
⁸ Department of Mathematics, Massachusetts Institute of Technology, Cambridge, MA.
⁹ Division of Neuro-Oncology, Translational Neuro-Oncology Laboratories, Department of Neurosciences, U.C. San Diego, Moores Cancer Center, La Jolla, CA.

^# Contributed equally.

Abstract

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

High-Throughput Nucleotide Sequencing*
Humans
Open Reading Frames / genetics*
Protein Binding
Protein Biosynthesis*
Protein Interaction Mapping / methods*
Protein Interaction Maps*
src-Family Kinases / metabolism

Substances

lyn protein-tyrosine kinase
src-Family Kinases

Abstract

Publication types

MeSH terms

Substances

Grants and funding