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Non-canonical amino acid labeling in proteomics and biotechnology
Metabolic labeling of proteins with non-canonical amino acids (ncAAs) provides unique bioorthogonal chemical groups during de novo synthesis by taking advantage of both endogenous and heterologous protein synthesis machineries. Labeled proteins can then be selectively conjugated to fluorophores, aff...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6529998/ https://www.ncbi.nlm.nih.gov/pubmed/31139251 http://dx.doi.org/10.1186/s13036-019-0166-3 |
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author | Saleh, Aya M. Wilding, Kristen M. Calve, Sarah Bundy, Bradley C. Kinzer-Ursem, Tamara L. |
author_facet | Saleh, Aya M. Wilding, Kristen M. Calve, Sarah Bundy, Bradley C. Kinzer-Ursem, Tamara L. |
author_sort | Saleh, Aya M. |
collection | PubMed |
description | Metabolic labeling of proteins with non-canonical amino acids (ncAAs) provides unique bioorthogonal chemical groups during de novo synthesis by taking advantage of both endogenous and heterologous protein synthesis machineries. Labeled proteins can then be selectively conjugated to fluorophores, affinity reagents, peptides, polymers, nanoparticles or surfaces for a wide variety of downstream applications in proteomics and biotechnology. In this review, we focus on techniques in which proteins are residue- and site-specifically labeled with ncAAs containing bioorthogonal handles. These ncAA-labeled proteins are: readily enriched from cells and tissues for identification via mass spectrometry-based proteomic analysis; selectively purified for downstream biotechnology applications; or labeled with fluorophores for in situ analysis. To facilitate the wider use of these techniques, we provide decision trees to help guide the design of future experiments. It is expected that the use of ncAA labeling will continue to expand into new application areas where spatial and temporal analysis of proteome dynamics and engineering new chemistries and new function into proteins are desired. |
format | Online Article Text |
id | pubmed-6529998 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-65299982019-05-28 Non-canonical amino acid labeling in proteomics and biotechnology Saleh, Aya M. Wilding, Kristen M. Calve, Sarah Bundy, Bradley C. Kinzer-Ursem, Tamara L. J Biol Eng Review Metabolic labeling of proteins with non-canonical amino acids (ncAAs) provides unique bioorthogonal chemical groups during de novo synthesis by taking advantage of both endogenous and heterologous protein synthesis machineries. Labeled proteins can then be selectively conjugated to fluorophores, affinity reagents, peptides, polymers, nanoparticles or surfaces for a wide variety of downstream applications in proteomics and biotechnology. In this review, we focus on techniques in which proteins are residue- and site-specifically labeled with ncAAs containing bioorthogonal handles. These ncAA-labeled proteins are: readily enriched from cells and tissues for identification via mass spectrometry-based proteomic analysis; selectively purified for downstream biotechnology applications; or labeled with fluorophores for in situ analysis. To facilitate the wider use of these techniques, we provide decision trees to help guide the design of future experiments. It is expected that the use of ncAA labeling will continue to expand into new application areas where spatial and temporal analysis of proteome dynamics and engineering new chemistries and new function into proteins are desired. BioMed Central 2019-05-22 /pmc/articles/PMC6529998/ /pubmed/31139251 http://dx.doi.org/10.1186/s13036-019-0166-3 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Review Saleh, Aya M. Wilding, Kristen M. Calve, Sarah Bundy, Bradley C. Kinzer-Ursem, Tamara L. Non-canonical amino acid labeling in proteomics and biotechnology |
title | Non-canonical amino acid labeling in proteomics and biotechnology |
title_full | Non-canonical amino acid labeling in proteomics and biotechnology |
title_fullStr | Non-canonical amino acid labeling in proteomics and biotechnology |
title_full_unstemmed | Non-canonical amino acid labeling in proteomics and biotechnology |
title_short | Non-canonical amino acid labeling in proteomics and biotechnology |
title_sort | non-canonical amino acid labeling in proteomics and biotechnology |
topic | Review |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6529998/ https://www.ncbi.nlm.nih.gov/pubmed/31139251 http://dx.doi.org/10.1186/s13036-019-0166-3 |
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