Eukaryotic cells are like tiny, highly organized cities, filled with many different compartments (organelles) that each have specific jobs. These compartments are surrounded by membranes and need to be carefully managed to function properly. A group of proteins called Rab GTPases act as traffic controllers, helping to organize and direct movement within the cell. However, we still don’t fully understand how these proteins create order in such a busy and complex environment.
So far, most research on Rab GTPases has either been performed in living cells or by studying them in very simplified experiments. However, neither approach is sufficient to reveal or explain their complex behavior in the cell. We now know that that Rab GTPases work as part of intricate biochemical networks that can self-organize—like a city that manages itself rather than needing constant outside control. To truly understand how they function, new experimental methods are needed.
In this project, we will take a “bottom-up” approach, rebuilding the Rab GTPase networks from scratch using purified components. This will allow us to see how they naturally organize themselves outside of a cell while still providing experimental access to uncover the underlying molecular interactions. We will also create artificial cell-like environments using microfabrication techniques to understand how these proteins respond to physical and chemical signals, similar to what happens in real cells. Finally, we will use cryo-electron microscopy techniques to look at the molecular structure of Rab GTPases and their partners.
By combining these approaches, this project will give us a better understanding of how Rab GTPases control the biochemical identities of intracellular membranes. This will also help us see how cells maintain their internal order, which is crucial for their survival and function.
