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Puromycin-associated nascent chain proteomics (PUNCH-P) for monitoring protein synthesis

PUNCH-P could be used to identify and quantify rapid proteome responses in disease states. The PUNCH-P process involves cell-free labeling of nascent proteins with biotinylated puromycin followed by a streptavidin-affinity purification step and analysis with liquid chromatography-tandem mass spectrometry. In human cell lines, PUNCH-P quantified mRNA translation products with accuracy comparable to that of two established proteomic approaches and was able to monitor cell cycle-specific fluctuations in mRNA translation for over 5,000 proteins. PUNCH-P also was used to generate a protein translation profile of the mouse brain from ex vivo samples. Next steps include evaluating PUNCH-P for diseases of the brain and for monitoring cancer progression.

SciBX 6(35); doi:10.1038/scibx.2013.971
Published online Sept. 12, 2013

Unpatented; licensing status not applicable

Aviner, R. et al. Genes Dev.; published online Aug. 9, 2013;
doi:10.1101/gad.219105.113
Contact: Orna Elroy-Stein, Tel Aviv University, Tel Aviv, Israel
e-mail:
ornaes@tauex.tau.ac.il

Contact: Tamar Geiger, same affiliation as above
e-mail:
geiger@post.tau.ac.il