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Engineered bacterial host for genetic encoding of physiologically stable protein nitration
Across scales, many biological phenomena, such as protein folding or bioadhesion and cohesion, rely on synergistic effects of different amino acid side chains at multiple positions in the protein sequence. These are often fine-tuned by post-translational modifications that introduce additional chemi...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9638147/ https://www.ncbi.nlm.nih.gov/pubmed/36353730 http://dx.doi.org/10.3389/fmolb.2022.992748 |
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author | Koch, Nikolaj G. Baumann, Tobias Nickling, Jessica H. Dziegielewski, Anna Budisa, Nediljko |
author_facet | Koch, Nikolaj G. Baumann, Tobias Nickling, Jessica H. Dziegielewski, Anna Budisa, Nediljko |
author_sort | Koch, Nikolaj G. |
collection | PubMed |
description | Across scales, many biological phenomena, such as protein folding or bioadhesion and cohesion, rely on synergistic effects of different amino acid side chains at multiple positions in the protein sequence. These are often fine-tuned by post-translational modifications that introduce additional chemical properties. Several PTMs can now be genetically encoded and precisely installed at single and multiple sites by genetic code expansion. Protein nitration is a PTM of particular interest because it has been associated with several diseases. However, even when these nitro groups are directly incorporated into proteins, they are often physiologically reduced during or shortly after protein production. We have solved this problem by using an engineered Escherichia coli host strain. Six genes that are associated with nitroreductase activity were removed from the genome in a simple and robust manner. The result is a bacterial expression host that can stably produce proteins and peptides containing nitro groups, especially when these are amenable to modification. To demonstrate the applicability of this strain, we used this host for several applications. One of these was the multisite incorporation of a photocaged 3,4-dihydroxyphenylalanine derivative into Elastin-Like Polypeptides. For this non-canonical amino acid and several other photocaged ncAAs, the nitro group is critical for photocleavability. Accordingly, our approach also enhances the production of biomolecules containing photocaged tyrosine in the form of ortho-nitrobenzyl-tyrosine. We envision our engineered host as an efficient tool for the production of custom designed proteins, peptides or biomaterials for various applications ranging from research in cell biology to large-scale production in biotechnology. |
format | Online Article Text |
id | pubmed-9638147 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-96381472022-11-08 Engineered bacterial host for genetic encoding of physiologically stable protein nitration Koch, Nikolaj G. Baumann, Tobias Nickling, Jessica H. Dziegielewski, Anna Budisa, Nediljko Front Mol Biosci Molecular Biosciences Across scales, many biological phenomena, such as protein folding or bioadhesion and cohesion, rely on synergistic effects of different amino acid side chains at multiple positions in the protein sequence. These are often fine-tuned by post-translational modifications that introduce additional chemical properties. Several PTMs can now be genetically encoded and precisely installed at single and multiple sites by genetic code expansion. Protein nitration is a PTM of particular interest because it has been associated with several diseases. However, even when these nitro groups are directly incorporated into proteins, they are often physiologically reduced during or shortly after protein production. We have solved this problem by using an engineered Escherichia coli host strain. Six genes that are associated with nitroreductase activity were removed from the genome in a simple and robust manner. The result is a bacterial expression host that can stably produce proteins and peptides containing nitro groups, especially when these are amenable to modification. To demonstrate the applicability of this strain, we used this host for several applications. One of these was the multisite incorporation of a photocaged 3,4-dihydroxyphenylalanine derivative into Elastin-Like Polypeptides. For this non-canonical amino acid and several other photocaged ncAAs, the nitro group is critical for photocleavability. Accordingly, our approach also enhances the production of biomolecules containing photocaged tyrosine in the form of ortho-nitrobenzyl-tyrosine. We envision our engineered host as an efficient tool for the production of custom designed proteins, peptides or biomaterials for various applications ranging from research in cell biology to large-scale production in biotechnology. Frontiers Media S.A. 2022-10-24 /pmc/articles/PMC9638147/ /pubmed/36353730 http://dx.doi.org/10.3389/fmolb.2022.992748 Text en Copyright © 2022 Koch, Baumann, Nickling, Dziegielewski and Budisa. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Molecular Biosciences Koch, Nikolaj G. Baumann, Tobias Nickling, Jessica H. Dziegielewski, Anna Budisa, Nediljko Engineered bacterial host for genetic encoding of physiologically stable protein nitration |
title | Engineered bacterial host for genetic encoding of physiologically stable protein nitration |
title_full | Engineered bacterial host for genetic encoding of physiologically stable protein nitration |
title_fullStr | Engineered bacterial host for genetic encoding of physiologically stable protein nitration |
title_full_unstemmed | Engineered bacterial host for genetic encoding of physiologically stable protein nitration |
title_short | Engineered bacterial host for genetic encoding of physiologically stable protein nitration |
title_sort | engineered bacterial host for genetic encoding of physiologically stable protein nitration |
topic | Molecular Biosciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9638147/ https://www.ncbi.nlm.nih.gov/pubmed/36353730 http://dx.doi.org/10.3389/fmolb.2022.992748 |
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