Analytical Ultracentrifuge

描述(由申请者提供)：路易斯维尔大学正在推动结构生物学和癌症研究，作为其核心使命的一部分。在JG布朗癌症中心和化学系的支持下，一些核心设施和重大合作项目正在建立中。目前这包括核磁共振光谱学、X射线结晶学、分子建模和蛋白质表达。为了能够对大分子及其在溶液状态下的相互作用进行功能分析，我们现在正在建立一个由一名资深科学家领导的生物物理核心。我们已经确定分析性超速离心法是这一核心的一个组成部分。任何功能分析的一个基本部分是确定分离的组分的物理状态(包括相对分子质量、形状、水合作用和均质性)，以及测量络合物的化学计量比和相互作用强度。由于参数对形状和尺寸的依赖，用其他非平衡方法很难做到这一点。分析超速离心法是为数不多的能够独立、严格地测定溶液中处于非变性状态的功能组分质量的技术之一，它还能提供有关均一性和摩擦性能的信息。此外，在考虑完全的核磁共振结构确定之前，通常有必要知道齐聚物或络合物的化学计量比，并且最好是对称性。分析超速离心机是一种理想的共享仪器，因为它可以很容易地容纳几个研究小组，他们的主要方法不是大分子的流体动力学，但他们需要它提供的具体数据，以进一步他们的研究和解决其他技术无法解决的问题。大分子的流体动力学分析可以被视为癌症中心整体生物物理学计划的一个组成部分。结构和生物物理技术的结合对于理解细胞内的调控过程以及在药物发现和开发过程中提供有关基本药物靶点的基本信息是至关重要的。有许多由美国国立卫生研究院资助的研究人员需要使用这种仪器，他们将是主要用户。还有许多其他群体需要不那么频繁的访问。所有研究人员都积极参与蛋白质表达和与疾病状态相关的复杂相互作用的分析。

DESCRIPTION (provided by applicant): The University of Louisville is promoting structural biology and cancer research as part of its core mission. Under the aegis of the JG Brown Cancer Center and the Department of Chemistry, a number of core facilities and major collaborative programs are being established. This currently includes NMR spectroscopy, X-ray crystallography, Molecular Modeling, and protein expression. To enable the functional analysis of macromolecules and their interactions in the solution state, we are now building a biophysics core to be headed by a senior scientist. We have identified analytical ultracentrifugation as an integral component of this core. An essential part of any functional analysis is the determination of the physical state of the isolated components (including molecular weight, shape, hydration and homogeneity), as well as measuring the stoichiometry of the complexes and the interaction strength. This is difficult to achieve by other nonequilibrium methods because of the dependent of the parameters on both shape and size. The analytical ultracentrifuge is one of the few techniques that can independently and rigorously determine mass of a functional component in a non-denaturing state in solution, as well as provide information about homogeneity and frictional properties. Furthermore, it is generally necessary to know the stoichiometry, and preferably the symmetry, of an oligomer or complex before a complete NMR structure determination can be contemplated. The analytical ultracentrifuge is an ideal shared instrument, as it can readily accommodate several research groups whose primary methodologies are not in the hydrodynamics of macromolecules, but who need the specific data it provides in order to further their research and solve problems that cannot be tackled by other techniques. Hydrodynamic analysis of macromolecules can be seen as an integral part of the overall biophysics program at the Cancer Center. The combination of structural and biophysical techniques is central to understanding regulatory processes inside cells, and for providing the basic information about basic drug targets during the drug discovery and development process. There are numerous NIH-funded investigators who need access to this instrumentation, and who would be major users. There are many other groups who will need less frequent access. All of the researchers are actively involved in protein expression and analysis of complex interactions related to disease states.