Correlative Cryo-electron Tomography Pipeline of Plasma Membrane Complexes

This cross-institute team calls on the unique expertise and skills of three labs and will develop tools to map the visual plasma membrane proteome. This project will build a new universal pipeline for solving protein and organelle structures at the plasma membrane using correlative cryogenic fluorescence and electron tomography. The team will streamline the preparation of thin mammalian cell plasma membranes in vitreous ice and develop probes for the precise localization of specific membrane-associated factors within electron tomograms. Using these probes, researchers will solve near-atomic structures of identified factors and their binding partners. The team will also design open modular freeware that accelerates correlative cryogenic fluorescence and electron tomography pipelines. With this information in mind, the team will benchmark tools across cell types, methods, and target proteins. These protocol and software developments will enable the biomedical community to solve the structures of a currently inaccessible set of molecular complexes that are central to human health and disease.