Collaborative Research: EAGER: Uncovering the role of Golgi organization on function

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Synthetic cell engineering has the potential to reveal insights into the fundamental chemical and physical processes that underlie the function of biological cells and organelles as well as advance the design of biologically-inspired devices and materials with applications ranging from medicine to biotechnology. Recently, it has become clear that the function of many proteins and lipids in biological cells depends on posttranslational modifications, covalent additions to the surface of proteins after their synthesis. A majority of these modifications are performed by the Golgi apparatus, a central membrane organelle in mammalian cells with a unique and dynamically changing structure. Yet neither the features of the Golgi that facilitate the synthesis of these highly complex chemical structures nor how those reactions are spatially and temporally controlled is fully understood. To better understand the regulatory mechanisms of the Golgi structure, this proposal will reconstitute chemical and physical features of the Golgi in an in vitro system. This proposal will develop new strategies to uncover how physical features of membranes influence an essential posttranslational modification, glycosylation. Specifically, it will recreate a model multi-step glycosylation modification of lipid carriers in vitro using both microfluidic and vesicle-based platforms and uncover the rules of membrane spatial and physical features required to recapitulate the posttranslational modification process. Successful completion of these studies will reveal design rules that will enable creation of other posttranslational modifications through re-organization of these reactions, including synthetic modifications. With these insights, it will be possible to extend these findings to influence the activity and function of modified proteins and lipids in order to impact biological processes. This project will bridge a variety of interdisciplinary techniques in membrane biophysics, membrane protein reconstitution, protein engineering, microfluidics, transport phenomena, and chemical kinetics necessary to realize the project goals. The knowledge developed here will benefit the biotechnology and pharmaceutical industries and have potential biomanufacturing applications in the design of therapeutic compounds. This project is jointly funded by the Cellular Dynamics and Function and Systems and Synthetic Biology Clusters, Division of Molecular and Cellular Biosciences, Directorate for Biological Sciences and by the Cellular and Biochemical Engineering Program, Division of Chemical, Bioengineering, Environmental and Transport Systems, Directorate for Engineering.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.