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High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain
During storage in the silk gland, the N-terminal domain (NT) of spider silk proteins (spidroins) keeps the aggregation-prone repetitive region in solution at extreme concentrations. We observe that NTs from different spidroins have co-evolved with their respective repeat region, and now use an NT th...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6959368/ https://www.ncbi.nlm.nih.gov/pubmed/31937841 http://dx.doi.org/10.1038/s41598-019-57143-x |
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author | Abelein, Axel Chen, Gefei Kitoka, Kristīne Aleksis, Rihards Oleskovs, Filips Sarr, Médoune Landreh, Michael Pahnke, Jens Nordling, Kerstin Kronqvist, Nina Jaudzems, Kristaps Rising, Anna Johansson, Jan Biverstål, Henrik |
author_facet | Abelein, Axel Chen, Gefei Kitoka, Kristīne Aleksis, Rihards Oleskovs, Filips Sarr, Médoune Landreh, Michael Pahnke, Jens Nordling, Kerstin Kronqvist, Nina Jaudzems, Kristaps Rising, Anna Johansson, Jan Biverstål, Henrik |
author_sort | Abelein, Axel |
collection | PubMed |
description | During storage in the silk gland, the N-terminal domain (NT) of spider silk proteins (spidroins) keeps the aggregation-prone repetitive region in solution at extreme concentrations. We observe that NTs from different spidroins have co-evolved with their respective repeat region, and now use an NT that is distantly related to previously used NTs, for efficient recombinant production of the amyloid-β peptide (Aβ) implicated in Alzheimer’s disease. A designed variant of NT from Nephila clavipes flagelliform spidroin, which in nature allows production and storage of β-hairpin repeat segments, gives exceptionally high yields of different human Aβ variants as a solubility tag. This tool enables efficient production of target peptides also in minimal medium and gives up to 10 times more isotope-labeled monomeric Aβ peptides per liter bacterial culture than previously reported. |
format | Online Article Text |
id | pubmed-6959368 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-69593682020-01-17 High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain Abelein, Axel Chen, Gefei Kitoka, Kristīne Aleksis, Rihards Oleskovs, Filips Sarr, Médoune Landreh, Michael Pahnke, Jens Nordling, Kerstin Kronqvist, Nina Jaudzems, Kristaps Rising, Anna Johansson, Jan Biverstål, Henrik Sci Rep Article During storage in the silk gland, the N-terminal domain (NT) of spider silk proteins (spidroins) keeps the aggregation-prone repetitive region in solution at extreme concentrations. We observe that NTs from different spidroins have co-evolved with their respective repeat region, and now use an NT that is distantly related to previously used NTs, for efficient recombinant production of the amyloid-β peptide (Aβ) implicated in Alzheimer’s disease. A designed variant of NT from Nephila clavipes flagelliform spidroin, which in nature allows production and storage of β-hairpin repeat segments, gives exceptionally high yields of different human Aβ variants as a solubility tag. This tool enables efficient production of target peptides also in minimal medium and gives up to 10 times more isotope-labeled monomeric Aβ peptides per liter bacterial culture than previously reported. Nature Publishing Group UK 2020-01-14 /pmc/articles/PMC6959368/ /pubmed/31937841 http://dx.doi.org/10.1038/s41598-019-57143-x Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Abelein, Axel Chen, Gefei Kitoka, Kristīne Aleksis, Rihards Oleskovs, Filips Sarr, Médoune Landreh, Michael Pahnke, Jens Nordling, Kerstin Kronqvist, Nina Jaudzems, Kristaps Rising, Anna Johansson, Jan Biverstål, Henrik High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain |
title | High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain |
title_full | High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain |
title_fullStr | High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain |
title_full_unstemmed | High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain |
title_short | High-yield Production of Amyloid-β Peptide Enabled by a Customized Spider Silk Domain |
title_sort | high-yield production of amyloid-β peptide enabled by a customized spider silk domain |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6959368/ https://www.ncbi.nlm.nih.gov/pubmed/31937841 http://dx.doi.org/10.1038/s41598-019-57143-x |
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