Differential refractometer and SAXS

伴随着结构生物学的快速发展，生物化学的功能和机制被定义为蛋白质相互作用中的分子作用力以及蛋白质的稳定性和折叠。对蛋白质结构的了解并不能确保对功能和生物活性的准确预测。为了充分表征蛋白质与底物、其他蛋白质或配体(如底物、核酸、小分子配体或离子)结合时的特定生物分子相互作用，分子理解蛋白质相互作用中的驱动力需要了解许多不同的参数。热力学提供了有关驱动生物分子相互作用的能量的信息。然后可以利用对催化、结合和稳定过程的这种理解来处理疾病途径背后的复杂过程，并制定抗击疾病的战略。这种策略最常见的例子是基于它们与特定酶或调节蛋白结合的可能性来设计新药。此外，正在设计的新药可以干扰或刺激人类和动物体内的代谢途径，以改善导致疾病症状的异常生化过程。申请人实验室的研究重点是了解与催化周转有关的亚基间信号、极端嗜热古菌蛋白的蛋白质稳定性、蛋白质-核酸识别以及叶绿素生物合成的关键酶GSAM的稳定性和结合特性。一个专注于蛋白质结构-功能关系研究的现代生物化学实验室结合了许多技术，将从蛋白质结构确定到目标系统的生化和生物物理表征的步骤整合在一起。该应用所需的设备是这一战略的组成部分，包括用于非常精确的蛋白质和配体浓度测定的差示折射仪，以及用于分析生物分子相互作用的热力学参数以研究蛋白质结合和蛋白质稳定性的分子细节的现有内部小角X射线散射(SAXS)设备的试管样品转换器。

Accompanying the rapid advances in structural biology is the definition of biochemical function and mechanism in terms of molecular forces in protein interactions and protein stability and folding. Knowledge of protein structure does not ensure accurate prediction of function and biological activity. To fully characterize specific biomolecular interactions in protein binding of substrates, other proteins or ligands such as substrates, nucleic acids, small molecule ligands or ions, a molecular understanding of driving forces in protein interactions requires knowledge of a number of different parameters. Thermodynamics provides information on the energetic forces that drive biomolecular interactions. This understanding of catalytic, binding and stability processes can then be exploited to tackle the complex processes underlying disease pathways and to devise strategies for combating diseases. The most common example of such strategies is the design of new drugs based on the likelihood of their binding to specific enzymes or regulatory proteins. In addition, new drugs are being designed that either interfere with or stimulate metabolic pathways within humans and animals to ameliorate aberrant biochemical processes causing disease symptoms. Research in the laboratory of the applicant is focused on understanding intersubunit signaling linked to catalytic turnover, protein stability of extremophilic archea proteins, protein-nucleic acid recognition, and stability and binding properties of GSAM, a key enzyme in chlorophyll biosynthesis. A modern biochemistry laboratory focused on protein study of structure-function relationships combines a number of technologies to integrate the steps from protein structure determination to biochemical and biophysical characterization of the target systems. The equipment requested in this application is integral to this strategy and includes a differential refractometer for very precise protein and ligand concentration determinations, and a Cuvette Sample Changer for an existing in-house Small-Angle X-Ray Scattering (SAXS) facility to analyze thermodynamic parameters of biomolecular interactions to study molecular details of protein binding and protein stability.