Recombinant Bolaamphiphilic Protein Engineering for Asymmetric Vesicle Formation toward Synthetic Cell Signal Transduction

This project addresses a fundamental challenge in synthetic biology: how to design cell-like systems that can sense their environment and process information in a controlled and programmable way. Living cells rely on asymmetric membranes and precisely organized proteins to convert external signals into internal responses, enabling essential functions such as communication, recognition, and adaptation. However, recreating these capabilities in synthetic systems remains a major scientific barrier. This research will develop a new class of fully protein-based vesicles that mimic these key cellular features, enabling directional sensing and programmable signal processing. By establishing fundamental principles that link molecular organization to biological function, the project promotes the progress of science and advances the frontiers of biotechnology—an area of strategic importance for national health, economic competitiveness, and innovation. The outcomes will enable new approaches in biosensing and bio-inspired materials. In addition, the project will provide interdisciplinary training for undergraduate and graduate students in protein engineering, biomaterials, and synthetic biology, while engaging K–12 students through hands-on modules and outreach programs. These activities will broaden participation in STEM, strengthen the future workforce, and enhance public understanding of emerging biotechnologies, thereby advancing the nation’s scientific enterprise and societal well-being. This research aims to engineer recombinant globular protein vesicles (GPVs) with controlled membrane asymmetry and integrated signal transduction capabilities. The project will pursue three objectives: (1) design and synthesize modular bolaamphiphilic fusion proteins that self-assemble into asymmetric vesicle membranes with defined protein orientation; (2) construct inward signal transduction pathways that convert ligand binding at the vesicle surface into lumenal biochemical outputs using mechanisms such as split enzyme reconstitution, protease cascades, and cell-free gene expression; and (3) quantitatively determine how protein orientation, spatial distribution, and lateral mobility within the membrane influence signaling efficiency. The approach integrates recombinant protein engineering, bioconjugation chemistry, advanced fluorescence imaging, cryo-electron microscopy, and small-angle scattering to characterize membrane structure and dynamics across multiple length scales. By correlating membrane architecture with functional outputs, the project will establish predictive design rules for programmable synthetic cells. These outcomes will advance synthetic biology and biomolecular engineering by providing foundational knowledge and enabling technologies for constructing adaptive, cell-like systems with tunable sensing and response behaviors. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria. NSF Award ID: 2540253 | Program: 01002627DB NSF RESEARCH & RELATED ACTIVIT | Principal Investigator: Yeongseon Jang | Institution: University of Florida, GAINESVILLE, FL | Award Amount: $692,207 View on NSF Award Search: https://www.nsf.gov/awardsearch/show-award/?AWD_ID=2540253 View on Research.gov: https://www.research.gov/awardapi-service/v1/awards/2540253.html