Crystallization Suite for the Structure Determination of Bacterial Cellulose Biosynthetic Proteins

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This grant will purchase a crystallography suite to support the applicant's research program in the structure-function characterization of bacterial cellulose biosynthetic proteins. Our functional studies are closely intertwined with our structural research as these areas complement each other to provide a detailed understanding of how enzyme activity and substrate binding relates to critical structural folds and/or motifs. The requested crystallography suite will advance our structural research by permitting the efficient generation and identification of X-ray diffraction quality crystals suitable for protein structure determination. Modern programmable incubators will allow for precise control over parameters needed for optimal crystal growth. An innovative stereomicroscope equipped with high magnification, a UV-light module and polarized light filters will greatly enhance our ability to unambiguously distinguish protein crystals from other artifacts in the field of view. The requested camera, software and automated features for the microscope enable specific control over image clarity, acquisition and processing which are important for reliable documentation and assessing crystal growth details related to reproducibility. Cryogenic storage dewars and crystal tools will facilitate the proper cryoprotection of protein crystals in order to minimize damage to crystals during transport and analysis while also providing a means for long-term storage. Thus, the requested instruments will provide a proper crystallization platform that enhances our ability to generate X-ray diffraction quality crystals suitable for protein structure determination. These results will provide a detailed understanding of the biosynthesis of bacterial cellulose and will lead to the discovery of important principles relating to polysaccharide synthesis and biofilm development across bacterial species. This crystallization suite will also serve to create a crystallography resource centre at Wilfrid Laurier University, thereby ensuring that trainees in the field of structure-function biochemistry are equipped with advanced skills/techniques that make them highly competitive upon entering Canada's workforce or pursuing higher education.