Protein structure determination facility upgrade

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Scope of research: All cellular functions are enabled by proteins. Understanding the structures of proteins and the architecture of protein complexes is the main gateway to understanding their functions. X-ray crystallography remains a gold standard method determining these macromolecular structures. Drs. Peter Davies and John Allingham lead research programs at Queen's University that have determined a broad spectrum of high-resolution protein structures using a home-source X-ray diffractometer that collects X-ray diffraction data from single proteins and protein complexes that have crystallized. Research programs to be supported: Dr. Davies is funded by NSERC to investigate the structures and mechanisms of proteins that mediate the interactions of bacteria with other cells or physical surfaces. Dr. Allingham is funded by NSERC to determine the structures and mechanisms of motor proteins that can change the structure of the actin and microtubule cytoskeletons, thereby affecting the movement and division of cells. Davies and Allingham also apply X-ray crystallography in drug discovery since ligands and compounds with high affinity can often be observed in the binding pockets of proteins. Need for the requested equipment: The X-ray crystallography laboratory at Queen's was constructed in 2008 and houses a Rigaku MicroMax007-HighFlux X-ray generator, an R-AXIS VI++ image-plate detector, and an X-Stream-2000 cryogenic cooling system. Normally, the X-Stream-2000 maintains crystal samples in a frozen, protected state during X-ray diffraction data collection by cooling them to -175 °C in an open flow nitrogen gas stream, but this machine is no longer functional nor serviceable. Without the ability to cool the protein crystals they are rapidly degraded by the X-ray beam. Thus, although the other components of the half million-dollar X-ray diffractometer are working well, we are completely unable to use it because of the broken cryostream. This application requests funding to purchase the Oxford Cryostream 1000 to replace this broken equipment. Training of HQP and sustainability: The X-ray crystallography laboratory was designed to allow experienced and novice crystallographers to complete a structural biology project from start to finish. Practical training and supervision in purifying target proteins, performing crystallization screening experiments, operating the diffractometer for crystal quality analysis and data collection, and solving protein structures is provided by Dr. Allingham, the X-ray lab director. Upgrades to this local research infrastructure is essential for these researchers to regain the ability to solve protein structures at Queen's and for maximizing their productivity during remote data diffraction data collections at synchrotron facilities. This equipment will also be critical for recruiting from the growing cohort of new scientists that are being trained in the fields of X-ray crystallography and cryo-electron microscopy.