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Controllable protein phase separation and modular recruitment to form responsive membraneless organelles

Many intrinsically disordered proteins self-assemble into liquid droplets that function as membraneless organelles. Because of their biological importance and ability to colocalize molecules at high concentrations, these protein compartments represent a compelling target for bio-inspired materials e...

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Autores principales: Schuster, Benjamin S., Reed, Ellen H., Parthasarathy, Ranganath, Jahnke, Craig N., Caldwell, Reese M., Bermudez, Jessica G., Ramage, Holly, Good, Matthew C., Hammer, Daniel A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6065366/
https://www.ncbi.nlm.nih.gov/pubmed/30061688
http://dx.doi.org/10.1038/s41467-018-05403-1
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author Schuster, Benjamin S.
Reed, Ellen H.
Parthasarathy, Ranganath
Jahnke, Craig N.
Caldwell, Reese M.
Bermudez, Jessica G.
Ramage, Holly
Good, Matthew C.
Hammer, Daniel A.
author_facet Schuster, Benjamin S.
Reed, Ellen H.
Parthasarathy, Ranganath
Jahnke, Craig N.
Caldwell, Reese M.
Bermudez, Jessica G.
Ramage, Holly
Good, Matthew C.
Hammer, Daniel A.
author_sort Schuster, Benjamin S.
collection PubMed
description Many intrinsically disordered proteins self-assemble into liquid droplets that function as membraneless organelles. Because of their biological importance and ability to colocalize molecules at high concentrations, these protein compartments represent a compelling target for bio-inspired materials engineering. Here we manipulated the intrinsically disordered, arginine/glycine-rich RGG domain from the P granule protein LAF-1 to generate synthetic membraneless organelles with controllable phase separation and cargo recruitment. First, we demonstrate enzymatically triggered droplet assembly and disassembly, whereby miscibility and RGG domain valency are tuned by protease activity. Second, we control droplet composition by selectively recruiting cargo molecules via protein interaction motifs. We then demonstrate protease-triggered controlled release of cargo. Droplet assembly and cargo recruitment are robust, occurring in cytoplasmic extracts and in living mammalian cells. This versatile system, which generates dynamic membraneless organelles with programmable phase behavior and composition, has important applications for compartmentalizing collections of proteins in engineered cells and protocells.
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spelling pubmed-60653662018-07-31 Controllable protein phase separation and modular recruitment to form responsive membraneless organelles Schuster, Benjamin S. Reed, Ellen H. Parthasarathy, Ranganath Jahnke, Craig N. Caldwell, Reese M. Bermudez, Jessica G. Ramage, Holly Good, Matthew C. Hammer, Daniel A. Nat Commun Article Many intrinsically disordered proteins self-assemble into liquid droplets that function as membraneless organelles. Because of their biological importance and ability to colocalize molecules at high concentrations, these protein compartments represent a compelling target for bio-inspired materials engineering. Here we manipulated the intrinsically disordered, arginine/glycine-rich RGG domain from the P granule protein LAF-1 to generate synthetic membraneless organelles with controllable phase separation and cargo recruitment. First, we demonstrate enzymatically triggered droplet assembly and disassembly, whereby miscibility and RGG domain valency are tuned by protease activity. Second, we control droplet composition by selectively recruiting cargo molecules via protein interaction motifs. We then demonstrate protease-triggered controlled release of cargo. Droplet assembly and cargo recruitment are robust, occurring in cytoplasmic extracts and in living mammalian cells. This versatile system, which generates dynamic membraneless organelles with programmable phase behavior and composition, has important applications for compartmentalizing collections of proteins in engineered cells and protocells. Nature Publishing Group UK 2018-07-30 /pmc/articles/PMC6065366/ /pubmed/30061688 http://dx.doi.org/10.1038/s41467-018-05403-1 Text en © The Author(s) 2018 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
Schuster, Benjamin S.
Reed, Ellen H.
Parthasarathy, Ranganath
Jahnke, Craig N.
Caldwell, Reese M.
Bermudez, Jessica G.
Ramage, Holly
Good, Matthew C.
Hammer, Daniel A.
Controllable protein phase separation and modular recruitment to form responsive membraneless organelles
title Controllable protein phase separation and modular recruitment to form responsive membraneless organelles
title_full Controllable protein phase separation and modular recruitment to form responsive membraneless organelles
title_fullStr Controllable protein phase separation and modular recruitment to form responsive membraneless organelles
title_full_unstemmed Controllable protein phase separation and modular recruitment to form responsive membraneless organelles
title_short Controllable protein phase separation and modular recruitment to form responsive membraneless organelles
title_sort controllable protein phase separation and modular recruitment to form responsive membraneless organelles
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6065366/
https://www.ncbi.nlm.nih.gov/pubmed/30061688
http://dx.doi.org/10.1038/s41467-018-05403-1
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