Dynamic Light Scattering, Calorimeter and Fluorometer for Structure/Function Studies of Macromolecules

明尼苏达大学的6个主要用户和4个次要用户正在申请研究酶、肌肉蛋白、载脂蛋白、细菌毒素和金属蛋白的结构和功能所需的仪器。这些仪器将用于为正在进行的结构分析提供热力学和动力学特性的重要联系。研究人员正在利用明尼苏达大学最近建立的结构生物学核心实验室的共享设施，通过诱变和蛋白质设计来研究生物功能。所有用户都将使用动态光散射仪器来分析正在设计和生产的蛋白质的聚集状态。了解蛋白质或蛋白质复合物的寡聚态对于核磁共振研究、x射线衍射晶体的制备以及各种光谱分析都是必不可少的。三个主要和三个次要用户将使用滴定量热计来确定蛋白质配体结合的能量学，这通常需要在结构研究中对突变或共价修饰的蛋白质进行功能表征。三名主要用户和三名次要用户将使用多频相位荧光计来分析蛋白质动力学、结构和蛋白质-蛋白质相互作用。这个特别的重点将放在使用诱变来设计蛋白质荧光探针的标记位点，提供特定位点的结构和动态信息，以补充x射线和核磁共振结构研究。

Six major and four minor users at the University of Minnesota are requesting instrumentation needed to study the structure and function of enzymes, muscle proteins, apolipoproteins, bacterial toxins, and metalloproteins. These instruments will be used to provide a vital linkage of thermodynamic and dynamic properties to ongoing structural analyses. The investigators are studying biological function through mutagenesis and protein design, using the shared facilities of a recently established Structural Biology Core Laboratory at the University of Minnesota. A dynamic light scattering instrument will be employed by all of the users to analyze the aggregation state of proteins that are being designed and produced. Understanding the oligomeric state of a protein or protein complex is essential to NMR studies, to the preparation of crystals for X-ray diffraction, and to a variety of spectroscopic analyses. A titration calorimeter will be used by three major and three minor users to determine the energetics of protein ligand binding, which is often required for the functional characterization of mutant or covalently modified proteins in structural studies. A multifrequency phase fluorometer will be used by three major and three minor users to analyze protein dynamics, structure, and protein-protein interactions. This particular emphasis will be placed on the use of mutagenesis to engineer labeling sites for fluorescent probes in proteins, providing site-specific structural and dynamic information to complement x-ray and NMR structural studies.