Understanding the rules of life in order to build synthetic cells that recapitulate processes of living organisms is a promising, emerging field that could ultimately define what it means to “be alive” and enable control of life in unprecedented ways that can benefit mankind. Achieving this knowledge will require understanding how to synthesize some of the basic biological building blocks of living cells. Of particular interest to this project is the Golgi apparatus, an organelle that precisely builds glycans for the posttranslational modification (PTM) of proteins and lipids to regulate cellular processes. 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. The overall objective of this project is to gain this critical understanding of this organelle, by deconstructing it into in vitro systems that allow testing of the constraints of modularity, sequence, and spatial arrangement of the enzymes that carry out glycosylation and drive this major PTM function of the Golgi. This team brings distinct and complementary expertise and is uniquely qualified to address these open research areas. Specific aims are to: (1) measure PTM efficiency as a function of enzyme spatial organization using patterned biomembrane devices, (2) measure how membrane biophysical features influence the spatial dynamics of model glycosylation enzymes and PTM efficiency on both 2-D planar bilayers and 3-D vesicular structures, and (3) integrate artificial Golgi vesicles with living systems. This project will create synthetic Golgi systems to define the relationships between enzyme spatial arrangement, membrane physical features, and efficiency of a model PTM process. It will also focus on elucidating the importance of spatial organization on final glycan structure and determining if modularity is a guiding principle in Golgi function that must be maintained in synthetic living systems.
|Effective start/end date||8/15/19 → 7/31/21|
- National Science Foundation (MCB-1935356)