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Engineering receptors in the secretory pathway for orthogonal signalling control
Synthetic receptors targeted to the secretory pathway often fail to exhibit the expected activity due to post-translational modifications (PTMs) and/or improper folding. Here, we engineered synthetic receptors that reside in the cytoplasm, inside the endoplasmic reticulum (ER), or on the plasma memb...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9708828/ https://www.ncbi.nlm.nih.gov/pubmed/36446786 http://dx.doi.org/10.1038/s41467-022-35161-0 |
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author | Mahameed, Mohamed Wang, Pengli Xue, Shuai Fussenegger, Martin |
author_facet | Mahameed, Mohamed Wang, Pengli Xue, Shuai Fussenegger, Martin |
author_sort | Mahameed, Mohamed |
collection | PubMed |
description | Synthetic receptors targeted to the secretory pathway often fail to exhibit the expected activity due to post-translational modifications (PTMs) and/or improper folding. Here, we engineered synthetic receptors that reside in the cytoplasm, inside the endoplasmic reticulum (ER), or on the plasma membrane through orientation adjustment of the receptor parts and by elimination of dysfunctional PTMs sites. The cytoplasmic receptors consist of split-TEVp domains that reconstitute an active protease through chemically-induced dimerization (CID) that is triggered by rapamycin, abscisic acid, or gibberellin. Inside the ER, however, some of these receptors were non-functional, but their activity was restored by mutagenesis of cysteine and asparagine, residues that are typically associated with PTMs. Finally, we engineered orthogonal chemically activated cell-surface receptors (OCARs) consisting of the Notch1 transmembrane domain fused to cytoplasmic tTA and extracellular CID domains. Mutagenesis of cysteine residues in CID domains afforded functional OCARs which enabled fine-tuning of orthogonal signalling in mammalian cells. |
format | Online Article Text |
id | pubmed-9708828 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-97088282022-12-01 Engineering receptors in the secretory pathway for orthogonal signalling control Mahameed, Mohamed Wang, Pengli Xue, Shuai Fussenegger, Martin Nat Commun Article Synthetic receptors targeted to the secretory pathway often fail to exhibit the expected activity due to post-translational modifications (PTMs) and/or improper folding. Here, we engineered synthetic receptors that reside in the cytoplasm, inside the endoplasmic reticulum (ER), or on the plasma membrane through orientation adjustment of the receptor parts and by elimination of dysfunctional PTMs sites. The cytoplasmic receptors consist of split-TEVp domains that reconstitute an active protease through chemically-induced dimerization (CID) that is triggered by rapamycin, abscisic acid, or gibberellin. Inside the ER, however, some of these receptors were non-functional, but their activity was restored by mutagenesis of cysteine and asparagine, residues that are typically associated with PTMs. Finally, we engineered orthogonal chemically activated cell-surface receptors (OCARs) consisting of the Notch1 transmembrane domain fused to cytoplasmic tTA and extracellular CID domains. Mutagenesis of cysteine residues in CID domains afforded functional OCARs which enabled fine-tuning of orthogonal signalling in mammalian cells. Nature Publishing Group UK 2022-11-29 /pmc/articles/PMC9708828/ /pubmed/36446786 http://dx.doi.org/10.1038/s41467-022-35161-0 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Mahameed, Mohamed Wang, Pengli Xue, Shuai Fussenegger, Martin Engineering receptors in the secretory pathway for orthogonal signalling control |
title | Engineering receptors in the secretory pathway for orthogonal signalling control |
title_full | Engineering receptors in the secretory pathway for orthogonal signalling control |
title_fullStr | Engineering receptors in the secretory pathway for orthogonal signalling control |
title_full_unstemmed | Engineering receptors in the secretory pathway for orthogonal signalling control |
title_short | Engineering receptors in the secretory pathway for orthogonal signalling control |
title_sort | engineering receptors in the secretory pathway for orthogonal signalling control |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9708828/ https://www.ncbi.nlm.nih.gov/pubmed/36446786 http://dx.doi.org/10.1038/s41467-022-35161-0 |
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