Macromolecular Crystallography Data Collection Instrumen

要求为最先进的大分子晶体学数据收集仪器提供资金。我们特别感兴趣的是购买一个集成系统，包括Rigaku RU- H3RHB x射线发生器，共聚焦光学系统（蓝色配置），反向phi轴，MSC R-Axis IV++区域探测器，x流低温系统和一台带有驱动仪器和处理衍射数据的软件的控制计算机。这个新系统将通过确保足够的x射线束时间用于晶体筛选/表征和完整的数据收集，直接并立即影响NIH资助的8个研究小组的项目。它将在停机期间补充我们已有9年历史的R-Axis II系统，该系统越来越需要对这种旧型号进行维护和维修，并适应我们对仪器日益增长的需求。在大分子晶体学数据收集的扩展能力是至关重要的，以追求以下项目：a)通过N- > O酰基移位进行自身蛋白水解，b) N端亲核水解酶，c) MspI作为一种新的结构类限制性内切酶，d)膜联蛋白晶体结构和膜相互作用，e)凝血中的金属依赖复合物，f)外排介导的四环素耐药性，g)表面活性剂蛋白的新抗氧化机制，h)磷酸化酶的结构和机制研究，i)醛缩酶的细胞功能解读，j)元裂变途径中的酶，k)光蛋白aequorin， 1)多重抗生素耐药抑制因子MarR， m)核小体组装伴侣，n)凋亡小体及其在程序性细胞死亡中的作用，o)脂蛋白结构和载脂蛋白构象，p)神经节苷脂gm1 -霍乱毒素，q) LDL受体-LDL复合物，r)胰岛素受体-胰岛素复合物，s)人载脂蛋白C-1。总的来说，这些项目将应用大分子晶体学来提高我们对生物分子复合物和细胞机器的功能和组装的理解。这些大分子组装在酶学、膜信号传导和染色质组装中发挥重要作用。此外，这些研究将深入了解糖蛋白功能以及LDL介导的脂质代谢的细胞过程，以及与动脉粥样硬化和AGU遗传疾病相关的正常和异常结构的形成。

Funds are requested for a state-of-the-art macromolecular crystallography data collection instrument. We are specifically interested in purchasing an integrated system with a Rigaku RU- H3RHB X-ray generator, a confocal optical system (blue configuration), an inverse phi axis, an MSC R-Axis IV++ area detector, an X-stream cryogenic system, and a control computer with software to drive the instrument and process diffraction data. This new system will directly and immediately impact NIH funded projects among 8 research groups by ensuring adequate X-ray beamtime for crystal screening/characterization and complete data collections. It will complement our 9 year old R-Axis II system during downtime that is increasingly required for maintenance and repairs of this older model, and accommodate our growing need for the instrument. The expanded capabilities in macromolecular crystallographic data collection are vital to pursue the following projects: a) autoproteolysis through N -> O acyl shift, b) N-terminal nucleophile hydrolase, c) MspI as a new structural class of restriction enzymes, d) annexin crystal structures and membrane interactions, e) metal dependent complexes in blood coagulation, f) efflux-mediated resistance to tetracyclines, g) novel antioxidant mechanisms of surfactant proteins, h) structural and mechanistic studies of phosphonatase, i) deciphering the cellular functions of aldolase, j) enzymes in the meta-fission pathway, k) photoprotein aequorin, 1) multiple antibiotic resistance repressor MarR, m) nucleosome assembly chaperones, n) the apoptosome and its role in programmed cell death, o) lipoprotein structure and apoprotein conformation, p) ganglioside GM1-cholera toxin, q) LDL receptor-LDL complex, r) insulin receptor-insulin complex, s) human apolipoprotein C-1. Overall, these projects will apply macromolecular crystallography to enhance our understanding of the function and assembly of biomolecular complexes and cellular machines. These macromolecular assemblies play important roles in enzymology, signalling through membranes, and chromatin assembly. Furthermore, these studies will provide insights into glycoprotein function as well as cellular processes of lipid metabolism mediated by LDL and the formation of both normal and aberrant structures with medical relevance to atherosclerosis and AGU genetic disease.