Journal
CELL CHEMICAL BIOLOGY
Volume 23, Issue 7, Pages 805-815Publisher
CELL PRESS
DOI: 10.1016/j.chembiol.2016.05.018
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Funding
- Medical Research Council, UK [MC_U105181009, MC_UP_A024_1008]
- King's College junior research fellowship
- MRC [MC_U105181009, MC_UP_A024_1008] Funding Source: UKRI
- Medical Research Council [MC_UP_A024_1008, MC_U105181009] Funding Source: researchfish
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Cell-specific proteomics in multicellular systems and whole animals is a promising approach to understand the differentiated functions of cells and tissues. Here, we extend our stochastic orthogonal recoding of translation (SORT) approach for the co-translational tagging of proteomes with a cyclopropene-containing amino acid in response to diverse codons in genetically targeted cells, and create a tetrazine-biotin probe containing a cleavable linker that offers a way to enrich and identify tagged proteins. We demonstrate that SORT with enrichment, SORT-E, efficiently recovers and enriches SORT tagged proteins and enables specific identification of enriched proteins via mass spectrometry, including low-abundance proteins. We show that tagging at distinct codons enriches overlapping, but distinct sets of proteins, suggesting that tagging at more than one codon enhances proteome coverage. Using SORT-E, we accomplish cell-specific proteomics in the fly. These results suggest that SORT-E will enable the definition of cell-specific proteomes in animals during development, disease progression, and learning and memory.
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