Analytical Ultracentrifuge

分析超速离心机

• 批准号: 7216484
• 负责人: John Charles Williams
• 金额: $ 28.42万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-16 至 2008-01-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7216484
• 关键词: Address; Area; Arrestin; Arrestins; Binding; Binding Proteins; Biological; CD8B1 gene; Complex; Core Facility; Data Collection; Data Quality; Detection; Dynein ATPase; Educational process of instructing; Equilibrium; Extinction (Psychology); G Protein-Coupled Receptor Signaling; Genetic; Glycoproteins; Graduate Education; Hand; Individual; Kimmel Cancer Center at the Thomas Jefferson University; Measurement; Measures; Membrane; Membrane Fusion; Methods; Molecular; Numbers; Optics; Peptide/MHC Complex; Peptides; Personal Satisfaction; Property; Protein Family; Proteins; Range; Refractive Indices; Regulation; Research Personnel; Roentgen Rays; Sampling; Screening procedure; Structure; Students; System; Techniques; Thermodynamics; Transfer RNA; Viral; dynactin; instrument; instrumentation; protein protein interaction; small molecule; structural biology

DESCRIPTION (provided by applicant): A critical number of investigators at the Kimmel Cancer Center of Thomas Jefferson University have a number of pending projects that require a quantitative method for measuring the quaternary structure of biological samples and their association constants. To this end, we propose to acquire the Beckman Optima XL-I analytical ultracentrifuge. This instrument is well suited for these projects and significantly extends the capabilities of our molecular interaction core facility. Specific areas that we will immediately address using this instrumentation are: 1) Thermodynamic characterization of multimeric dynein and dynactin complexes in relation to the regulation of dynein activity and cargo binding; 2) To study the oligomerization state and hydrodynamic properties of peptide segments derived from viral glycoproteins involved in membrane fusion; 3) tRNA-protein complexes and the mechanism of decoding genetic information; 4) Characterization of Protein-Protein Interaction Antagonists; 5) Characterize conformational changes induced by activation of arrestins, a family of proteins that regulate G protein-coupled receptor signaling; 6) Characterization of the equilibrium binding constants and the stochiometry of the ectodomains of the TCR, the pMHC and the CD8. In addition, the instrument will enhance two additional core facilities (X-ray and small molecule screening facilities) as well as graduate education (PR613, a graduate course on structural biology, is structured such that students apply each technique taught to an actual sample). The analytical ultracentrifuge of choice is the Beckman Optima XL-I. This instrument permits detection using either interference optics or UV/visible optics. The interference mode is sensitive to the refractive index, and thus, does not require the analyte to have a natural chromaphor. Thus, this will permit measurement of samples with low molecular extinction coefficients, small molecule antagonist and membrane bound proteins. The interference optics also provides rapid data collection useful for producing high quality data for sedmentation velocity analysis. On the other hand, the ability to measure absorbance over a large range (190 to 800 nM) allows simultaneous measurement of individual components in a multicomponent system.

描述（由申请人提供）：Thomas Jefferson大学Kimmel癌症中心的一些研究人员有一些待完成的项目，这些项目需要一种定量方法来测量生物样品的四元结构及其结合常数。为此，我们建议购买贝克曼Optima XL-I分析超离心机。该仪器非常适合这些项目，并显着扩展了我们的分子相互作用核心设施的能力。我们将使用该仪器立即解决的具体领域是：1)与动力蛋白活性和货物结合调节相关的多聚动力蛋白和动力蛋白复合物的热力学表征；2)研究参与膜融合的病毒糖蛋白衍生肽段的寡聚状态和流体动力学性质；3) trna -蛋白复合物及其解码遗传信息的机制；4)蛋白-蛋白相互作用拮抗剂的表征；5)表征由抑制蛋白（调控G蛋白偶联受体信号的蛋白家族）激活引起的构象变化；6) TCR、pMHC和CD8的平衡结合常数和外结构域的化学性质。此外，该仪器将加强两个额外的核心设施（x射线和小分子筛选设施）以及研究生教育（PR613，一门结构生物学研究生课程，旨在使学生将所教授的每种技术应用于实际样品）。选择的分析超离心机是贝克曼Optima XL-I。该仪器允许使用干涉光学或紫外/可见光光学进行检测。干涉模式是敏感的折射率，因此，不需要分析物具有自然色相。因此，这将允许测量样品具有低分子消光系数，小分子拮抗剂和膜结合蛋白。干涉光学还提供了快速的数据收集，有助于为沉降速度分析提供高质量的数据。另一方面，在大范围内（190至800 nM）测量吸光度的能力允许在多组分系统中同时测量单个组分。