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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...

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Autores principales: Elliott, Thomas S., Bianco, Ambra, Townsley, Fiona M., Fried, Stephen D., Chin, Jason W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cell Press 2016
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.
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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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