Structural Biology Shared Resource

项目概要：结构生物学共享资源 氨基酸侧链三维排列的原子图像给出了所需的原子细节 为了可视化酶的活性位点，查看转录因子的DNA结合位点，并查看 信号分子的蛋白质间相互作用。函数可以通过确定 X射线晶体学的原子结构。修饰大分子的功能是开发 无副作用的特定疗法。在没有晶体的情况下， 复合物、单个蛋白质及其在配体结合后的总体构象变化可以被确定 使用小角X射线散射（SAXS）。SBF（结构生物学设施）是许多年前发起的， 通过内布拉斯加州研究计划（NRI）和联合国大学负责研究的副校长的资助， （VCR）。它被癌症中心成员广泛使用，在支持癌症治疗科学方面发挥着重要作用。 癌症中心导致其指定为癌症中心核心设施开始于2008年。在2013年 到2015年，从NRI和VCR获得了1，741，452美元用于升级。SBF有四个主要的 实验室服务： 1.蛋白表达和纯化（PrEP）。 2.晶体筛选和生长（CSG）。 3. X射线（小角X射线散射（SAXS）和单晶衍射）。 5.核磁共振（NMR）数据收集。 PrEP实验室提供高质量的纯化重组蛋白样品，可进行同位素标记 用于NMR数据收集，准备结晶或其他实验。CSG实验室包括国家- 最先进的微型机器人结晶仪器。SAXS，单晶X射线 晶体学和NMR用于结构测定。SBF有两名联合董事（一名与 一个在蛋白质化学和X射线方法方面有专长，另一个在核磁共振方面有专长）， 具有结构测定经验。两名经理，一名技术员和一名博士后 用于开发蛋白质纯化和结晶方案，维护高科技仪器， 确保收集/处理最佳数据，并培训用户/学生。

PROJECT SUMMARY: STRUCTURAL BIOLOGY SHARED RESOURCE Atomic images of the arrangement of amino acid side chains in three dimensions give the atomic detail needed to visualize the active sites of enzymes, see the DNA-binding sites of transcription factors, and view the protein-protein interactions of signaling molecules. Function can be understood through the determination of atomic structures by X-ray crystallography. Modification of the function of macromolecules is key to developing specific therapies without side effects. In the absence of crystals, the molecular envelopes of macromolecular complexes, individual proteins, and their gross conformational changes upon ligand binding can be determined using small-angle X-ray scattering (SAXS). The SBF (Structural Biology Facility) was initiated many years ago, through funding from the Nebraska Research Initiative (NRI) and the UNMC Vice Chancellor for Research (VCR). Its expanding use by Cancer Center members and its important role in supporting the science of the Cancer Center led to its designation as a Cancer Center core facility beginning in 2008. In the years 2013 through 2015, $1,741,452 was obtained from the NRI and VCR for upgrades. The SBF has four main laboratory services: 1. Protein expression and purification (PrEP). 2. Crystal screening and growth (CSG). 3. X-ray (Small-angle X-ray scattering (SAXS) and single crystal diffraction). 5. Nuclear magnetic resonance (NMR) data collection. The PrEP laboratory provides high-quality purified recombinant protein samples that can be isotopically labeled for NMR data collection, ready for crystallization, or other experiments. The CSG laboratory includes state-of- the-art robotic crystallization instruments with microscale capabilities. SAXS, single-crystal X-ray crystallography, and NMR are used for structure determination. The SBF has two co-directors (one with expertise in protein chemistry and X-ray methods and the other with expertise in NMR) who share the decision making and have experience in structure determination. Two managers, a technician, and one postdoc are employed to develop protein purification and crystallization protocols, maintain the high-tech instrumentation, ensure that the best data is collected/processed, and train users/students.