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Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine
Site-specific incorporation of un-natural amino acids (UNAA) is a powerful approach to engineer and understand protein function. Site-specific incorporation of UNAAs is achieved through repurposing the amber codon (UAG) as a sense codon for the UNAA, using a tRNA(CUA) that base pairs with an UAG cod...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6566996/ https://www.ncbi.nlm.nih.gov/pubmed/31130675 http://dx.doi.org/10.3390/ijms20102577 |
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author | Syed, Junetha Palani, Saravanan Clarke, Scott T. Asad, Zainab Bottrill, Andrew R. Jones, Alexandra M.E. Sampath, Karuna Balasubramanian, Mohan K. |
author_facet | Syed, Junetha Palani, Saravanan Clarke, Scott T. Asad, Zainab Bottrill, Andrew R. Jones, Alexandra M.E. Sampath, Karuna Balasubramanian, Mohan K. |
author_sort | Syed, Junetha |
collection | PubMed |
description | Site-specific incorporation of un-natural amino acids (UNAA) is a powerful approach to engineer and understand protein function. Site-specific incorporation of UNAAs is achieved through repurposing the amber codon (UAG) as a sense codon for the UNAA, using a tRNA(CUA) that base pairs with an UAG codon in the mRNA and an orthogonal amino-acyl tRNA synthetase (aaRS) that charges the tRNA(CUA) with the UNAA. Here, we report an expansion of the zebrafish genetic code to incorporate the UNAAs, azido-lysine (AzK), bicyclononyne-lysine (BCNK), and diazirine-lysine (AbK) into green fluorescent protein (GFP) and glutathione-s-transferase (GST). We also present proteomic evidence for UNAA incorporation into GFP. Our work sets the stage for the use of AzK, BCNK, and AbK introduction into proteins as a means to investigate and engineer their function in zebrafish. |
format | Online Article Text |
id | pubmed-6566996 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-65669962019-06-17 Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine Syed, Junetha Palani, Saravanan Clarke, Scott T. Asad, Zainab Bottrill, Andrew R. Jones, Alexandra M.E. Sampath, Karuna Balasubramanian, Mohan K. Int J Mol Sci Communication Site-specific incorporation of un-natural amino acids (UNAA) is a powerful approach to engineer and understand protein function. Site-specific incorporation of UNAAs is achieved through repurposing the amber codon (UAG) as a sense codon for the UNAA, using a tRNA(CUA) that base pairs with an UAG codon in the mRNA and an orthogonal amino-acyl tRNA synthetase (aaRS) that charges the tRNA(CUA) with the UNAA. Here, we report an expansion of the zebrafish genetic code to incorporate the UNAAs, azido-lysine (AzK), bicyclononyne-lysine (BCNK), and diazirine-lysine (AbK) into green fluorescent protein (GFP) and glutathione-s-transferase (GST). We also present proteomic evidence for UNAA incorporation into GFP. Our work sets the stage for the use of AzK, BCNK, and AbK introduction into proteins as a means to investigate and engineer their function in zebrafish. MDPI 2019-05-26 /pmc/articles/PMC6566996/ /pubmed/31130675 http://dx.doi.org/10.3390/ijms20102577 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Communication Syed, Junetha Palani, Saravanan Clarke, Scott T. Asad, Zainab Bottrill, Andrew R. Jones, Alexandra M.E. Sampath, Karuna Balasubramanian, Mohan K. Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine |
title | Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine |
title_full | Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine |
title_fullStr | Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine |
title_full_unstemmed | Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine |
title_short | Expanding the Zebrafish Genetic Code through Site-Specific Introduction of Azido-lysine, Bicyclononyne-lysine, and Diazirine-lysine |
title_sort | expanding the zebrafish genetic code through site-specific introduction of azido-lysine, bicyclononyne-lysine, and diazirine-lysine |
topic | Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6566996/ https://www.ncbi.nlm.nih.gov/pubmed/31130675 http://dx.doi.org/10.3390/ijms20102577 |
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