Imaging protein crystals for identification of submicron nucleation and growth optimization

蛋白质三维结构的研究为了解生物过程的基本机制提供了宝贵的见解。X射线晶体学提供了一种以原子分辨率解决蛋白质结构的奇妙方法。本提案中的三位共同申请人迫切需要这些知识，以在膜蛋白的结构生物学（Martin Audet）、半胱天冬酶的酶学（Jean-Bernard Denault）和G蛋白细胞生物学（克莉丝汀Lavoie）方面开展令人兴奋的发现计划。正如提案中所详述的那样，共同申请人将极大地受益于对参与调节他们所研究的分子系统的蛋白质和辅因子的高分辨率描述。Martin Audet是一名早期职业研究员，其研究项目专注于溶质载体转运蛋白的基于X射线晶体学的结构生物学。Jean-Bernard Denault研究半胱天冬酶激活、调节和底物识别的机制。最后，克莉丝汀Lavoie专注于非传统的细胞角色的异源三聚体的GalphaS蛋白，特别是泛素化的作用上的结构功能的GalphaS。为了支持他们的NSERC研究计划和HQP培训，共同申请人需要获得设备，使他们能够成像和筛选大量结晶条件，以使用X射线晶体学方法确定其系统中重要蛋白质的高分辨率结构。用于识别蛋白质晶体的成像机器人使用多种检测策略来解决高复杂性（成千上万种条件）的结晶筛选，并以及时和可重复的方式轻松检索和分类蛋白质晶体。目前舍布鲁克缺乏这种设备，严重阻碍了这些发现计划的进展。结晶板成像的自动化以及所需设备适用于蛋白质晶体的检测方法的可用性将解锁共同申请人的研究计划，使我们能够与其他学术和工业合作伙伴开始新的合作，并最终为HQP提供一种仪器，该仪器将提高他们在X射线蛋白质晶体学中最先进的成像技术的技能。这种专业知识在加拿大是罕见的，精通这一领域的HQP在行业中的需求很高。所要求的仪器将为培训HQP奠定坚实的基础，这些HQP将构成未来的劳动力，以在加拿大科学工业和学术界传播结构生物学和X射线晶体学的杠杆作用。

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.