SPLIT: Stable Protein Coacervation Using a Light Induced Transition
Year: 2020
Type: article
Abstract: Protein coacervates serve as hubs to concentrate and sequester proteins and nucleotides and thus function as membraneless organelles to manipulate cell physiology. We have engineered a coacervating protein to create tunable, synthetic membraneless organelles that assemble in response to a single pulse of light. Coacervation is driven by the intrinsically disordered RGG domain from the protein LAF-1, and opto-responsiveness is coded by the protein PhoCl, which cleaves in response to 405 nm light. We developed a fusion protein containing a solubilizing maltose-binding protein domain, PhoCl, and two copies of the RGG domain. Several seconds of illumination at 405 nm is sufficient to cleave PhoCl, removing the solubilization domain and enabling RGG-driven coacervation within minutes in cellular-sized water-in-oil emulsions. An optimized version of this system displayed light-induced coacervation in Saccharomyces cerevisiae . The methods described here provide novel strategies for inducing protein phase separation using light.
Source: ACS Synthetic Biology
Language: English
Topic: RNA Research and Splicing
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: G1051554911, G155958390, Materials Research Science and Engineering Center, NEW; DESIGNING SMART, RESPONSIVE COMMUNICATING MICROCAPSULES FROM POLYMERSOMES; PI - HAMMER, DANIEL A., TAS::89 0331::TAS RECOVERY RECOVERY ACT: BLOCK GRANT FOR ARRA FUNDING. NEW AWARD FOR TUSTIN, CA, Programmable Synthetic Organelles Built from Disordered Proteins for Cellular Engineering, Engineering protein vesicles for drug delivery applications
OpenAlex ID: W3008604026