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

拟议的研究涉及单线态氧和其他衍生自由基的生产后，可见光照射的蛋白质属于家庭的绿色荧光蛋白。在这个提议中，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定量。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理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引起的损伤的表征和定量。最后，单线态氧的生产由不同的蛋白质在可见光照射后将被量化，并将被用来氧化模型系统（脂质），但也收到很少或根本没有attention.The大范围的荧光蛋白将使我们能够开发一类新的光敏剂，可能会发现应用在细胞中的氧化应激与亚细胞空间分辨率的产生。

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.