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

点击翻译按钮获取中文摘要

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.