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Spectrofluorimeter instrument for the investigation of fluorescent protein photobleaching mediated by singlet oxygen

用于研究单线态氧介导的荧光蛋白光漂白的荧光分光光度计仪器

基本信息

批准号：
359664-2008
负责人：
Heyne, Belinda
金额：
$ 3.25万
依托单位：
University of Calgary
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=365437
关键词：
Spectrofluorimeter instrument investigation fluorescent protein

项目摘要

The proposed research deals with the production of singlet oxygen and other derived radicals upon visible irradiation of proteins belonging to the family of the green fluorescent protein. Within this proposal, we will dedicate considerable effort to build synthetic compounds that mimic the different protein chromophores. The ability of these synthetic compounds to produce singlet oxygen will be followed and quantified in homogeneous and heterogeneous systems using state of the art spectroscopic techniques. Since these compounds are composed of well-known singlet oxygen inhibitors, a part of the proposed research will be dedicated to understanding and quantifying the interaction of singlet oxygen with the synthetic compounds mimicking the fluorescent protein chromophores. Another aspect of our work will also involve the characterization and quantification of the damage induced by singlet oxygen on the proteins themselves. Finally, the production of singlet oxygen by different proteins upon visible irradiation will be quantified and will be use to oxidize a model system (lipids) but also proteins that have received little or no attention.The large range of fluorescent proteins will allow us to develop a new class of photosensitizers that may find application in cells in the generation of oxidative stress with subcellular spatial resolution.

拟议的研究涉及绿色荧光蛋白家族的蛋白质在可见光照射下产生单线态氧和其他衍生自由基的问题。在这项提议中，我们将投入相当大的努力来构建模拟不同蛋白质发色团的合成化合物。这些合成化合物产生单线态氧的能力将在均相和非均相系统中使用最先进的光谱技术进行跟踪和量化。由于这些化合物由众所周知的单线态氧抑制剂组成，因此拟议研究的一部分将致力于了解和量化单线态氧与模拟荧光蛋白发色团的合成化合物的相互作用。我们工作的另一个方面也将涉及单线态氧对蛋白质本身造成的损伤的表征和量化。最后，不同蛋白质在可见光照射下产生的单线态氧将被量化，并将被用来氧化一个模型系统(脂类)，但也将被用来氧化那些很少或没有受到关注的蛋白质。大范围的荧光蛋白质将使我们能够开发一类新的光敏剂，可能在细胞中以亚细胞空间分辨率产生氧化应激。