Calorimetry Work Station

八名在蛋白质相互作用是重要组成部分的领域工作的BRI科学家提议购买一个由不同扫描量热(DSC)和等温滴定量热(ITC)仪器组成的量热工作站。该工作站将使他们能够确定生物大分子内部和之间的能量贡献，这是理解结构和功能之间关系的关键。研究人员目前的实验方法包括各种技术，包括X射线结晶学、荧光、超速离心法、CD和其他光谱方法以及免疫学和其他生物活性测量。测量蛋白质稳定性和蛋白质相互作用细节的量热方法将提供更定量的、由于可以获得的热力学参数，对这些研究中涉及的相互作用的更基本的描述。因此，决定结构的相互作用将与功能相关。BRI的量热工作站需要补充个别研究计划中详细说明的肌肉蛋白质、伴侣和免疫学物质的其他结构和功能研究，通常将用于：(I)表征蛋白质及其修饰的稳定性。这将有助于设计适合于功能和结晶学研究的突变体。(Ii)测量蛋白质结构域的热力学稳定性。DSC比CD更有用，因为结构域不需要太多的二级结构，而且这些系统中存在的多个跃迁更容易分析。(3)测量结合常数并表征结合平衡的热力学。ITC避免了荧光标记的不可避免的干扰，并且比结合的ELISA法更定量。

Eight BBRI scientists working in fields in which protein interactions are an important component, propose to purchase a Calorimetry Work Station consisting of different scanning calorimetry (DSC) and isothermal titration calorimetry (ITC) instrumentation. The work station will enable them to determine the energetic contributions within and between biological macromolecules which is key to understanding the relationship between structure and function. The current experimental approaches of the investigators involve a variety of techniques including X-ray crystallography, fluorescence, ultracentrifugation, CD and other spectroscopic methods as well as immunological and other measurements of biological activity. Calorimetric methods for measuring the details of protein stability and protein interactions would provide more quantitative, and because of the thermodynamic parameters obtainable, a more basic description of the interactions involved in these studies. Thus, interactions that determine structure will be related to function. The Calorimetry Work Station in BBRI is needed to complement other structural and functional studies with muscle proteins, chaperones and immunologicals as detailed in the individual research plans and generally will be used to: (i) characterize stability of proteins and their modifications. This will help in the design of suitable mutants for functional and crystallographic studies. (ii) measure thermodynamic stability of protein domains. DSC is more useful than CD because the domains need not have much secondary structure, and multiple transitions which are present in these systems are easier to analyze. (iii) measure binding constants and characterize the thermodynamics of binding equilibria. ITC avoids the unavoidable perturbation of fluorescence labels and is more quantitatively than ELISA assays of binding.