Calorimetry Work Station

量热工作站

• 批准号: 6440411
• 负责人: SHERWIN LEHRER
• 金额: $ 15.1万
• 依托单位: BOSTON BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2003-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6440411
• 关键词: biomedical equipment purchase; calorimetry; computers

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.

八位在蛋白质相互作用是重要组成部分领域工作的 BBRI 科学家提议购买由不同扫描量热法 (DSC) 和等温滴定量热法 (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涉及多种技术，包括X射线晶体学、荧光、超速离心、CD和其他光谱方法以及免疫学和其他生物活性测量。用于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多定量，并且由于可以获得热力学参数，因此可以对这些研究中涉及的相互作用提供更基本的描述。因此，决定结构的相互作用将与功能相关。 BBRI 的量热工作站需要补充其他肌肉蛋白、分子伴侣和免疫学的结构和功能研究，如个人研究计划中详述，通常用于：(i) 表征蛋白质及其修饰的稳定性。这将有助于设计适合功能和晶体学研究的突变体。 (ii) 测量蛋白质结构域的热力学稳定性。 DSC 比 CD 更有用，因为域不需要太多二级结构，并且这些系统中存在的多个转变更容易分析。 (iii) 测量结合常数并表征结合平衡的热力学。 ITC 避免了不可避免的荧光标记干扰，并且比 ELISA 结合测定更定量。