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Tagging and Enriching Proteins Enables Cell-Specific Proteomics
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 cyclopro...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cell Press
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4959846/ https://www.ncbi.nlm.nih.gov/pubmed/27447048 http://dx.doi.org/10.1016/j.chembiol.2016.05.018 |
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author | Elliott, Thomas S. Bianco, Ambra Townsley, Fiona M. Fried, Stephen D. Chin, Jason W. |
author_facet | Elliott, Thomas S. Bianco, Ambra Townsley, Fiona M. Fried, Stephen D. Chin, Jason W. |
author_sort | Elliott, Thomas S. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4959846 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Cell Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-49598462016-08-01 Tagging and Enriching Proteins Enables Cell-Specific Proteomics Elliott, Thomas S. Bianco, Ambra Townsley, Fiona M. Fried, Stephen D. Chin, Jason W. Cell Chem Biol Article 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. Cell Press 2016-07-21 /pmc/articles/PMC4959846/ /pubmed/27447048 http://dx.doi.org/10.1016/j.chembiol.2016.05.018 Text en © 2016 MRC Laboratory of Molecular Biology http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Elliott, Thomas S. Bianco, Ambra Townsley, Fiona M. Fried, Stephen D. Chin, Jason W. Tagging and Enriching Proteins Enables Cell-Specific Proteomics |
title | Tagging and Enriching Proteins Enables Cell-Specific Proteomics |
title_full | Tagging and Enriching Proteins Enables Cell-Specific Proteomics |
title_fullStr | Tagging and Enriching Proteins Enables Cell-Specific Proteomics |
title_full_unstemmed | Tagging and Enriching Proteins Enables Cell-Specific Proteomics |
title_short | Tagging and Enriching Proteins Enables Cell-Specific Proteomics |
title_sort | tagging and enriching proteins enables cell-specific proteomics |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4959846/ https://www.ncbi.nlm.nih.gov/pubmed/27447048 http://dx.doi.org/10.1016/j.chembiol.2016.05.018 |
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