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Intracellular phase separation of globular proteins facilitated by short cationic peptides
Phase separation provides intracellular organization and underlies a variety of cellular processes. These biomolecular condensates exhibit distinct physical and material properties. Current strategies for engineering condensate formation include using intrinsically disordered domains and altering pr...
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/PMC9780332/ https://www.ncbi.nlm.nih.gov/pubmed/36550144 http://dx.doi.org/10.1038/s41467-022-35529-2 |
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author | Yeong, Vivian Wang, Jou-wen Horn, Justin M. Obermeyer, Allie C. |
author_facet | Yeong, Vivian Wang, Jou-wen Horn, Justin M. Obermeyer, Allie C. |
author_sort | Yeong, Vivian |
collection | PubMed |
description | Phase separation provides intracellular organization and underlies a variety of cellular processes. These biomolecular condensates exhibit distinct physical and material properties. Current strategies for engineering condensate formation include using intrinsically disordered domains and altering protein surface charge by chemical supercharging or site-specific mutagenesis. We propose adding to this toolbox designer peptide tags that provide several potential advantages for engineering protein phase separation in bacteria. Herein, we demonstrate the use of short cationic peptide tags for sequestration of proteins of interest into bacterial condensates and provide a foundational study for their development as tools for condensate engineering. Using a panel of GFP variants, we demonstrate how cationic tag and globular domain charge contribute to intracellular phase separation in E. coli and observe that the tag can affect condensate disassembly at a given net charge near the phase separation boundary. We showcase the broad applicability of these tags by appending them onto enzymes and demonstrating that the sequestered enzymes remain catalytically active. |
format | Online Article Text |
id | pubmed-9780332 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-97803322022-12-24 Intracellular phase separation of globular proteins facilitated by short cationic peptides Yeong, Vivian Wang, Jou-wen Horn, Justin M. Obermeyer, Allie C. Nat Commun Article Phase separation provides intracellular organization and underlies a variety of cellular processes. These biomolecular condensates exhibit distinct physical and material properties. Current strategies for engineering condensate formation include using intrinsically disordered domains and altering protein surface charge by chemical supercharging or site-specific mutagenesis. We propose adding to this toolbox designer peptide tags that provide several potential advantages for engineering protein phase separation in bacteria. Herein, we demonstrate the use of short cationic peptide tags for sequestration of proteins of interest into bacterial condensates and provide a foundational study for their development as tools for condensate engineering. Using a panel of GFP variants, we demonstrate how cationic tag and globular domain charge contribute to intracellular phase separation in E. coli and observe that the tag can affect condensate disassembly at a given net charge near the phase separation boundary. We showcase the broad applicability of these tags by appending them onto enzymes and demonstrating that the sequestered enzymes remain catalytically active. Nature Publishing Group UK 2022-12-22 /pmc/articles/PMC9780332/ /pubmed/36550144 http://dx.doi.org/10.1038/s41467-022-35529-2 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 Yeong, Vivian Wang, Jou-wen Horn, Justin M. Obermeyer, Allie C. Intracellular phase separation of globular proteins facilitated by short cationic peptides |
title | Intracellular phase separation of globular proteins facilitated by short cationic peptides |
title_full | Intracellular phase separation of globular proteins facilitated by short cationic peptides |
title_fullStr | Intracellular phase separation of globular proteins facilitated by short cationic peptides |
title_full_unstemmed | Intracellular phase separation of globular proteins facilitated by short cationic peptides |
title_short | Intracellular phase separation of globular proteins facilitated by short cationic peptides |
title_sort | intracellular phase separation of globular proteins facilitated by short cationic peptides |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9780332/ https://www.ncbi.nlm.nih.gov/pubmed/36550144 http://dx.doi.org/10.1038/s41467-022-35529-2 |
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