Structural Biology Shared Resource

结构生物学共享资源

基本信息

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
项目类别：
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

项目摘要

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氨基酸侧链三维排列的原子图像给出了所需的原子细节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-蛋白质相互作用。函数可以通过确定来理解 X 射线晶体学的原子结构。大分子功能的修饰是开发的关键具体疗法无副作用。在没有晶体的情况下，大分子的分子包膜可以确定复合物、单个蛋白质及其配体结合时的总体构象变化使用小角 X 射线散射 (SAXS)。 SBF（结构生物学设施）成立于多年前，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）。癌症中心成员不断扩大其使用范围，并在支持癌症科学方面发挥重要作用癌症中心于 2008 年开始被指定为癌症中心核心设施。2013 年截至 2015 年，从 NRI 和 VCR 获得了 1,741,452 美元用于升级。 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，为结晶或其他实验做好准备。 CSG 实验室包括最先进的具有微型能力的最先进的机器人结晶仪器。 SAXS，单晶 X 射线晶体学和NMR用于结构测定。 SBF 有两名联合主任（其中一名是具有蛋白质化学和 X 射线方法方面的专业知识，另一位具有 NMR 方面的专业知识），共同做出决定制作并有结构测定的经验。两名经理、一名技术人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/处理最好的数据，并培训用户/学生。