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Artificial DnaJ Protein for protein production and conformational diseases
For secreted proteins, proper protein folding is essential not only for biological function but also for secretion itself. Proteins with folding problems are trapped in the endoplasmic reticulum (ER) and are eventually degraded in the cytoplasm. In this study, we exploited co-expression of an artifi...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5561034/ https://www.ncbi.nlm.nih.gov/pubmed/28819167 http://dx.doi.org/10.1038/s41598-017-09067-7 |
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author | Hishiya, Akinori Koya, Keizo |
author_facet | Hishiya, Akinori Koya, Keizo |
author_sort | Hishiya, Akinori |
collection | PubMed |
description | For secreted proteins, proper protein folding is essential not only for biological function but also for secretion itself. Proteins with folding problems are trapped in the endoplasmic reticulum (ER) and are eventually degraded in the cytoplasm. In this study, we exploited co-expression of an artificial fusion protein, based on the sequence of a DnaJ protein, which could interact as co-chaperones in the Hsp70-based protein-folding system, with target recombinant secreted proteins to enhance their production and secretion. The J-domain sequence or a fragment thereof was conjugated to a target protein–binding domain that was capable of binding to a portion of the target-protein sequence. Production of many of the target proteins was significantly upregulated when co-expressed with the J-domain fusion protein. Surprisingly, the enhancement of secretion was observed even when the J-domain had a mutation in the HPD motif, which is necessary for J-protein–Hsp70 interactions, suggesting the phenomenon observed is independent on functional J-protein–Hsp70 interactions. This technology has great potential for not only enhancing the production of recombinant proteins, but also to treat conformational diseases such as cystic fibrosis, and Alpha-1 antitrypsin deficiency. |
format | Online Article Text |
id | pubmed-5561034 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-55610342017-08-18 Artificial DnaJ Protein for protein production and conformational diseases Hishiya, Akinori Koya, Keizo Sci Rep Article For secreted proteins, proper protein folding is essential not only for biological function but also for secretion itself. Proteins with folding problems are trapped in the endoplasmic reticulum (ER) and are eventually degraded in the cytoplasm. In this study, we exploited co-expression of an artificial fusion protein, based on the sequence of a DnaJ protein, which could interact as co-chaperones in the Hsp70-based protein-folding system, with target recombinant secreted proteins to enhance their production and secretion. The J-domain sequence or a fragment thereof was conjugated to a target protein–binding domain that was capable of binding to a portion of the target-protein sequence. Production of many of the target proteins was significantly upregulated when co-expressed with the J-domain fusion protein. Surprisingly, the enhancement of secretion was observed even when the J-domain had a mutation in the HPD motif, which is necessary for J-protein–Hsp70 interactions, suggesting the phenomenon observed is independent on functional J-protein–Hsp70 interactions. This technology has great potential for not only enhancing the production of recombinant proteins, but also to treat conformational diseases such as cystic fibrosis, and Alpha-1 antitrypsin deficiency. Nature Publishing Group UK 2017-08-17 /pmc/articles/PMC5561034/ /pubmed/28819167 http://dx.doi.org/10.1038/s41598-017-09067-7 Text en © The Author(s) 2017 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 Hishiya, Akinori Koya, Keizo Artificial DnaJ Protein for protein production and conformational diseases |
title | Artificial DnaJ Protein for protein production and conformational diseases |
title_full | Artificial DnaJ Protein for protein production and conformational diseases |
title_fullStr | Artificial DnaJ Protein for protein production and conformational diseases |
title_full_unstemmed | Artificial DnaJ Protein for protein production and conformational diseases |
title_short | Artificial DnaJ Protein for protein production and conformational diseases |
title_sort | artificial dnaj protein for protein production and conformational diseases |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5561034/ https://www.ncbi.nlm.nih.gov/pubmed/28819167 http://dx.doi.org/10.1038/s41598-017-09067-7 |
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