Controllable protein phase separation and modular recruitment to form responsive membraneless organelles
Abstract
Abstract 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.
- Publication Date:
- Research Org.:
- Univ. of Pennsylvania, Philadelphia, PA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1461590
- Alternate Identifier(s):
- OSTI ID: 1511484
- Grant/Contract Number:
- SC0007063
- Resource Type:
- Published Article
- Journal Name:
- Nature Communications
- Additional Journal Information:
- Journal Name: Nature Communications Journal Volume: 9 Journal Issue: 1; Journal ID: ISSN 2041-1723
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Schuster, Benjamin S., Reed, Ellen H., Parthasarathy, Ranganath, Jahnke, Craig N., Caldwell, Reese M., Bermudez, Jessica G., Ramage, Holly, Good, Matthew C., and Hammer, Daniel A. Controllable protein phase separation and modular recruitment to form responsive membraneless organelles. United Kingdom: N. p., 2018.
Web. doi:10.1038/s41467-018-05403-1.
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. United Kingdom. https://doi.org/10.1038/s41467-018-05403-1
Schuster, Benjamin S., Reed, Ellen H., Parthasarathy, Ranganath, Jahnke, Craig N., Caldwell, Reese M., Bermudez, Jessica G., Ramage, Holly, Good, Matthew C., and Hammer, Daniel A. Mon .
"Controllable protein phase separation and modular recruitment to form responsive membraneless organelles". United Kingdom. https://doi.org/10.1038/s41467-018-05403-1.
@article{osti_1461590,
title = {Controllable protein phase separation and modular recruitment to form responsive membraneless organelles},
author = {Schuster, Benjamin S. and Reed, Ellen H. and Parthasarathy, Ranganath and Jahnke, Craig N. and Caldwell, Reese M. and Bermudez, Jessica G. and Ramage, Holly and Good, Matthew C. and Hammer, Daniel A.},
abstractNote = {Abstract 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.},
doi = {10.1038/s41467-018-05403-1},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United Kingdom},
year = {Mon Jul 30 00:00:00 EDT 2018},
month = {Mon Jul 30 00:00:00 EDT 2018}
}
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https://doi.org/10.1038/s41467-018-05403-1
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Cited by: 176 works
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Figures / Tables:
Fig. 1: Phase separation of RGG-based IDP constructs. a Illustration of four-cell C. elegans embryo. LAF-1 is present in P granules, which contribute to germline specification. b Domain organization of LAF-1, RGG, RGG-RGG, and RGG-RGG-RGG. c Representative turbidity measurements show temperaturedependent phase behavior of RGG, RGG-RGG, and RGG-RGG-RGG at proteinmore »
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