Tagging and Enriching Proteins Enables Cell-Specific Proteomics
| Autor: | Stephen D. Fried, Jason W. Chin, Ambra Bianco, Fiona M. Townsley, Thomas S. Elliott |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
Proteomics
0301 basic medicine Clinical Biochemistry Biotin Tetrazoles Computational biology Biology 010402 general chemistry 01 natural sciences Biochemistry Mass Spectrometry 03 medical and health sciences Drug Discovery Animals Drosophila Proteins Amino Acids Molecular Biology Pharmacology Genetics chemistry.chemical_classification Molecular Structure Translation (biology) Genetic code 0104 chemical sciences 3. Good health Transport protein Amino acid Protein Transport Multicellular organism Drosophila melanogaster 030104 developmental biology chemistry Genetic Code Molecular Probes Proteome Molecular Medicine Female Molecular probe Azo Compounds |
| Zdroj: | Cell Chemical Biology. 23(7):805-815 |
| ISSN: | 2451-9456 |
| DOI: | 10.1016/j.chembiol.2016.05.018 |
| Popis: | SummaryCell-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. |
| Databáze: | OpenAIRE |
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