Analytical Ultracentrifuge

描述（由申请人提供）：路易斯维尔大学正在促进结构生物学和癌症研究，作为其核心使命的一部分。在JG Brown癌症中心和化学系的支持下，正在建立一些核心设施和主要合作项目。这目前包括核磁共振光谱，x射线晶体学，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，我们正在建立一个由资深科学家领导的生物物理核心。我们已经确定了分析超离心作为这个核心的一个组成部分。任何功能分析的一个重要部分是确定分离组分的物理状态（包括分子量、形状、水合作用和均匀性），以及测量配合物的化学计量和相互作用强度。这是难以实现的其他非平衡方法，因为参数依赖于形状和尺寸。分析式超离心机是为数不多的能够独立而严格地测定溶液中处于非变性状态的功能组分的质量的技术之一，同时也能提供有关均匀性和摩擦特性的信息。此外，在可以考虑完整的核磁共振结构测定之前，通常有必要知道低聚物或复合物的化学计量学，最好是对称性。分析式超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，这些研究小组的主要方法不是大分子的流体动力学，而是需要它提供的特定数据来进一步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看作是癌症中心整体生物物理学项目的一个组成部分。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至关重要。有许多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较少的访问。所有的研究人员都积极参与与疾病状态相关的复杂相互作用的蛋白质表达和分析。

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.