Structural Biology Shared Resource

结构生物学共享资源

• 批准号: 10491817
• 负责人: Gloria Borgstahl
• 金额: $ 12.8万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-05 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10491817
• 关键词: 3-Dimensional; Active Sites; Amino Acids; Area; Binding Sites; Biological; Cancer Center; Cancer Center Support Grant; Cancer Science; Core Facility; Crystallization; DNA Binding; Data; Data Collection; Disease; Ensure; Enzymes; Fee-for-Service Plans; Funding; Growth; Image; Individual; Isotope Labeling; Knowledge; Laboratories; Ligand Binding; Macromolecular Complexes; Malignant Neoplasms; Methods; Modification; Molecular; Molecular Conformation; Nebraska; Nuclear Magnetic Resonance; Postdoctoral Fellow; Process; Protein Chemistry; Proteins; Protocols documentation; Recombinant Proteins; Research; Research Personnel; Resource Sharing; Robotics; Roentgen Rays; Role; Sampling; Services; Side; Signaling Molecule; Structure; Students; Techniques; Training; Training and Education; Work; X-Ray Crystallography; experience; experimental study; instrument; instrumentation; macromolecule; member; novel therapeutics; protein expression; protein protein interaction; protein purification; screening; shared decision making; side effect; structural biology; transcription factor

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.

项目摘要：结构生物学共享资源 在三维中氨基酸侧链排列的原子图像给出了所需的原子细节 要可视化酶的活性位点，请参阅转录因子的DNA结合位点，并查看 信号分子的蛋白质蛋白相互作用。可以通过确定来理解功能 X射线晶体学的原子结构。修改大分子的功能是开发的关键 没有副作用的特定疗法。在没有晶体的情况下，大分子的分子信封 可以确定配合物，单个蛋白质及其在配体结合上的构象变化。 使用小角度X射线散射（SAXS）。 SBF（结构生物学设施）是多年前启动的 通过内布拉斯加州研究计划（NRI）和UNMC研究副校长的资助 （录像机）。它不断扩大癌症中心成员的使用及其在支持科学方面的重要作用 癌症中心从2008年开始将其称为癌症中心核心设施。2013年 到2015年，从NRI和VCR获得了1,741,452美元的升级。 SBF有四个主要 实验室服务： 1。蛋白质表达和纯化（PREP）。 2。晶体筛查和生长（CSG）。 3。X射线（小角度X射线散射（SAXS）和单晶衍射）。 5。核磁共振（NMR）数据收集。 预备实验室提供高质量纯化的重组蛋白样品，可以在同位素上标记 对于NMR数据收集，准备结晶或其他实验。 CSG实验室包括 具有显微镜功能的艺术机器人结晶仪器。 SAXS，单晶X射线 晶体学和NMR用于结构测定。 SBF有两个联合导演（一个 分享决定 制定并具有结构确定的经验。两名经理，一名技术人员和一个博士后是 用于开发蛋白质纯化和结晶方案，维持高科技仪器， 确保收集/处理最佳数据，并培训用户/学生。