Structural Biology Shared Resource

项目摘要：结构生物学共享资源 氨基酸侧链排列的三维原子图像提供了所需的原子细节 要可视化酶的活性部位，请查看转录因子的DNA结合部位，并查看 信号分子的蛋白质-蛋白质相互作用。函数可以通过确定 用X-射线结晶学研究原子结构。大分子的功能修饰是发展的关键 无副作用的特效疗法。在没有晶体的情况下，大分子的分子包膜 可以确定复合体、单个蛋白质及其在配基结合时的总构象变化 使用小角X射线散射(SAXS)。SBF(结构生物学设施)是多年前启动的， 由内布拉斯加州研究倡议(NRI)和UNMC负责研究的副校长资助 (VCR)。它在癌症中心成员中的广泛应用及其在支持癌症科学中的重要作用 癌症中心使其从2008年开始被指定为癌症中心的核心设施。2013年 到2015年，从NRI和VCR获得了1,741,452美元的升级资金。SBF有四个主要部分 实验室服务： 1.蛋白质表达与纯化(PrEP)。 2.晶体筛选与生长(CSG)。 3.X射线(小角X射线散射和单晶衍射)。 5.核磁共振数据采集。 PrEP实验室提供高质量的纯化重组蛋白样本，可以进行同位素标记 用于核磁共振数据收集，准备结晶或进行其他实验。CSG实验室包括最新的 具有微尺度能力的最先进的机器人结晶仪器。单晶X射线小角X射线 用结晶学、核磁共振等方法进行了结构测定。SBF有两名联席董事(一名与 一个是蛋白质化学和X射线方法方面的专业知识，另一个是核磁共振方面的专业知识)，他们分享决定 制作，并有结构测定经验。两名经理、一名技术员和一名博士后 被用来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.