Circular dichroism spectrometer for analysis of protein structure and function

用于分析蛋白质结构和功能的圆二色光谱仪

• 批准号: 358903-2008
• 负责人: Hill, Bruce
• 金额: $ 6.27万
• 依托单位: Queen's University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2007
• 资助国家: 加拿大
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=364813
• 关键词: Circular; dichroism; spectrometer; analysis; protein

The functional status of proteins is critically dependent on their structure and most proteins go through a series of structural changes in the course of their functional cycle. Protein characterization involves being able to correlate the protein of interest's structural status and its functional state. Circular dichroism spectroscopy provides a powerful and relatively simple tool to assess a protein for its folded state and whether or not it has undergone a structural change upon ligand binding, enzyme turnover, or interaction with a protein binding partner.     Circular dichroism (i.e., CD) spectroscopy measures the inherent optical activity of any molecular center that absorbs light (i.e., chromophore) and has some inherent structural asymmetry. Protein molecules possess chromophores across the UV-visible spectrum. In the short ultraviolet the peptide linkage itself absorbs light, in the near UV aromatic residues such as tryptophan absorb light and there are prosthetic groups such as heme centers that absorb light in the visible and near infrared regions. Each of these chromophores may give rise to CD, and the nature of the signal is sensitive to the environment in which the chromophore resides. For example, the CD signal of a peptide bond is modified when it is in a secondary structure such as an a-helix and it is different when in a ß-sheet, or if the protein is unstructured. CD will be used to distinguish different conformational states of proteins of interest to our research groups. Understanding the evolution of a protein's structure over its functional cycle is a critical component to being able to predict, control and regulate a protein's activity.       The equipment requested here will provide an opportunity for the research personnel in our laboratories (students, technicians and post-doctoral fellows) to gain valuable training with one of the most widely used approaches in modern protein structure studies. Improving the training of young Canadians researchers in these areas is an important element in the development of our fundamental science and technology base.

蛋白质的功能状态主要取决于其结构，大多数蛋白质在其功能周期中经历一系列结构变化。蛋白质表征涉及能够将感兴趣的蛋白质的结构状态与其功能状态相关联。圆二色光谱提供了一个强大的和相对简单的工具来评估蛋白质的折叠状态，以及它是否经历了配体结合，酶周转，或与蛋白质结合伴侣的相互作用后的结构变化。 圆二色性（即，CD）光谱测量吸收光的任何分子中心的固有光学活性（即，发色团）并且具有一些固有的结构不对称性。蛋白质分子在紫外-可见光谱范围内具有发色团。在短紫外线下，肽键本身吸收光，在近紫外线下，色氨酸等芳香族残基吸收光，并且存在血红素中心等辅基，吸收可见光和近红外区域的光。这些发色团中的每一个都可以引起CD，并且信号的性质对发色团所在的环境敏感。例如，当肽键处于二级结构如α-螺旋中时，其CD信号被修饰，而当处于β-折叠中时，或者如果蛋白质是非结构化的，则其CD信号不同。CD将用于区分我们研究小组感兴趣的蛋白质的不同构象状态。了解蛋白质结构在其功能周期中的演变是能够预测，控制和调节蛋白质活性的关键组成部分。 这里要求的设备将为我们实验室的研究人员（学生，技术人员和博士后研究员）提供一个机会，以获得现代蛋白质结构研究中最广泛使用的方法之一的宝贵培训。改进对加拿大青年研究人员在这些领域的培训，是发展我国基本科学和技术基础的一个重要因素。