Crystallization Suite for the Structure Determination of Bacterial Cellulose Biosynthetic Proteins

这笔赠款将购买一个晶体学套件，以支持申请人在细菌纤维素生物合成蛋白质的结构-功能表征方面的研究计划。我们的功能研究与我们的结构研究密切相关，因为这些领域相互补充，以详细了解酶活性和底物结合与关键结构折叠和/或基序的关系。所要求的晶体学套件将通过允许有效生成和鉴定适合于蛋白质结构测定的X射线衍射质量晶体来推进我们的结构研究。现代可编程培养器将允许精确控制优化晶体生长所需的参数。一个创新的立体显微镜配备了高放大倍率，紫外光模块和偏振光过滤器将大大提高我们的能力，明确区分蛋白质晶体从其他文物的视野。显微镜所需的摄像头、软件和自动化功能可实现对图像清晰度、采集和处理的特定控制，这对于可靠记录和评估与再现性相关的晶体生长细节至关重要。低温储存杜瓦瓶和晶体工具将促进蛋白质晶体的适当低温保护，以最大限度地减少运输和分析过程中对晶体的损坏，同时还提供长期储存的手段。因此，所要求的仪器将提供一个适当的结晶平台，提高我们产生适合蛋白质结构测定的X射线衍射质量晶体的能力。这些结果将提供一个详细的了解细菌纤维素的生物合成，并将导致发现的重要原则，多糖合成和生物膜的发展跨越细菌物种。该结晶套件还将用于在Wilfrid Laurier大学创建晶体学资源中心，从而确保结构-功能生物化学领域的受训人员具备先进的技能/技术，使他们在进入加拿大劳动力市场或接受高等教育时具有高度竞争力。

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.