Imaging protein crystals for identification of submicron nucleation and growth optimization

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The study of the three-dimensional structure of proteins provides invaluable insight into the fundamental mechanisms of biological processes. X-ray crystallography provides a fantastic means to solve protein structures at atomic resolution. The three co-applicants in this proposal have an urgent need for such knowledge to pursue exciting Discovery Programs in the structural biology of membrane proteins (Martin Audet), enzymology of caspases (Jean-Bernard Denault), and G-protein cell biology (Christine Lavoie). As detailed in the proposal, the co-applicants would greatly benefit from a high-resolution description of proteins and cofactors involved in regulating the molecular systems they studied. Martin Audet is an early career researcher with a research program that focuses on X-ray crystallography-based structural biology of solute carrier transporters. Jean-Bernard Denault studies mechanisms of caspase activation, regulation, and substrate recognition. Finally, Christine Lavoie focuses on the nonconventional cellular roles of the heterotrimeric GalphaS protein, specifically the role of ubiquitination on the structure function of GalphaS. To support their NSERC research programs and HQP training, the co-applicants require access to equipment that would allow them to image and screen a large number of crystallization conditions to determine high-resolution structures of important proteins in their systems using an X-ray crystallography approach. Imager robots for the identification of protein crystals use multiple detection strategies to address crystallization screens of high complexity (dozens of thousands of conditions) and allow easy retrieval and classification of protein crystals in a timely and reproducible manner. The current lack of such equipment in Sherbrooke severely impedes advances in these Discovery Programs. Automatization of the imaging of the crystallization plates and the availability of detection methods adapted for protein crystals by the requested equipment will unlock the research program of the co-applicants, allow us to start new collaborations with other academic and industrial partners, and finally provide HQPs with an instrument that will elevate their skill set with state-of-the-art imaging technology in X-ray protein crystallography. This expertise is rare in Canada and HQPs that are proficient in this field is in high demand in the industry. The requested instrument will enable a solid ground to train HQPs that will constitute a future workforce to propagate the leverage of structural biology and X-ray crystallography in the Canadian scientific industry and academia.