Mechanism and Dynamics of the YidC Insertase in Membrane Protein Insertion

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This project examines how the bio-machine named YidC inserts and folds proteins into the membrane bilayer. Protein insertion into membranes is one of the most critical processes in biology, and understanding the mechanisms by which proteins are transported across or integrated into membranes has enormous potential impact for understanding cellular and organismal structure, function and health. The project will yield, as yet undiscovered, fundamental information on the function of YidC, which is key to understanding how biological membranes are assembled, maintain their identity, and support many functions such as signaling and transport. The resulting new knowledge on biogenesis of membrane proteins could also impact human health-related research in the future since membrane proteins make up more than 50% of the known drug targets. The project will also provide opportunities for training students in a multidisciplinary research environment involving molecular biology, biochemical and biophysical approaches. The goal of the research is to determine how YidC functions to insert proteins into the cell membrane. YidC is novel as a membrane insertase as it contains a hydrophilic cavity that is crucial for its insertase and chaperone function. YidC can function on its own or cooperate with the Sec machinery to insert proteins. The project utilizes fluorescence spectroscopy, electron paramagnetic resonance spin labeling, and single molecule approaches to examine the dynamics and structural relationships of membrane insertion events. The research specifically aims to determine the mechanism of action and dynamics of YidC in membrane protein insertion, elucidate the interactions in the YidC/Sec machinery complex, and identify features of membrane protein substrates that determine selection of YidC and Sec translocase pathways in E. coli. The project will advance the field of membrane protein insertion in bacteria and will impact understanding of similar pathways in human cells.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.