Fluorimeter for biophysical characterizations

用于生物物理表征的荧光计

• 批准号: 375336-2009
• 负责人: Pawelek, Peter
• 金额: $ 2.87万
• 依托单位: Concordia 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: 2008
• 资助国家: 加拿大
• 起止时间: 2008-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=412136
• 关键词: Fluorimeter; biophysical; characterizations

Fluorescence spectroscopy (flourimetry) is a technique that we use to better understand how the conformation of biological macromolecules (protein, DNA, RNA) can determine function. By measuring light emitted from a flourophore (a molecule that fluoresces upon excitation by light of a given wavelength) we can obtain useful information on the local chemical environments either within or on the surface of a protein. A fluorophore can either be intrinsic (e.g., an amino acid that is part of a protein) or extrinsic (e.g., a fluorescent dye attached to a protein). We use fluorimetry to report on the extent to which a protein-associated fluorophore is buried or quenched, since the amount of light emitted from such a fluorophore can increase or decrease dramatically as a result of protein conformational changes. Fluorescence measurements are performed with an instrument called a fluorescence spectrophotometer (fluorimeter). We need a fluorimeter to characterize the structural nature of our proteins as well as to gain an understanding of how they interact with each other. In the past year, our Department's single research-dedicated fluorimeter completely ceased functioning after 11 years and many repairs. We urgently require a modern fluorimeter with polarization and temperature control capabilities to advance our research programs. The use of fluorescence polarization is central to our fluorimetry needs given its power in probing protein conformational changes and protein-protein interactions. Also, many of us require accurate temperature control in order to see how changes in temperature affect fluorescence properties of our proteins or protein-fluorophore conjugates.

荧光光谱(荧光法)是一种我们用来更好地了解生物大分子(蛋白质、DNA、RNA)的构象如何决定功能的技术。通过测量荧光团(一种在特定波长的光激发下发出荧光的分子)发出的光，我们可以获得关于蛋白质内部或表面局部化学环境的有用信息。荧光团既可以是内在的(例如，作为蛋白质一部分的氨基酸)，也可以是外在的(例如，附着在蛋白质上的荧光染料)。我们使用荧光法来报告与蛋白质相关的荧光团被埋藏或猝灭的程度，因为这种荧光团发出的光量可能会因为蛋白质构象的变化而急剧增加或减少。荧光测量是用一种叫做荧光分光光度计(荧光计)的仪器进行的。我们需要一个荧光仪来表征我们蛋白质的结构性质，以及了解它们是如何相互作用的。在过去的一年里，我司单一的研究专用荧光仪在经过11年的多次维修后完全停止工作。我们迫切需要一种具有偏振和温度控制能力的现代荧光仪来推进我们的研究计划。由于荧光偏振在探测蛋白质构象变化和蛋白质-蛋白质相互作用方面的能力，它的使用对于我们的荧光测量需求是核心的。此外，我们中的许多人都需要精确的温度控制，以便了解温度的变化如何影响我们的蛋白质或蛋白质-荧光团偶合物的荧光性质。