Time-resolved spectroscopic characterization of blue-light photo-receptors

我们分析了衣藻蓝光受体Phot1的lov1结构域的结构和光化学过程。我们建议分析被认为发生在LOV1 （fmn型）三重态下的电子转移过程，并研究Phot1中信号状态的后续形成。将采用高度跨学科的方法，结合分子生物学、生物化学、光谱学、晶体学和理论。其次，我们计划研究含有所谓BLUF结构域的感觉蓝光受体作为发色团（FAD-type）。我们将表达在紫色细菌r.s sphaeroides中参与光合蛋白基因调控的AppA-Protein和鞭毛虫e.g arilis中作为光调节腺苷酸环化酶的PAC-a/b。我们将表征光激活的酶活性，并试图确定两种蛋白质在黑暗形式和光激活状态下的结构。我们将通过时间和波长分辨紫外-可见光谱和荧光光谱研究主要的光过程，特别是在ns时间范围内。光谱和动力学信息将与结构数据相结合，详细描述参与信号状态形成的物理和光化学反应。

We have analysed the structure and the photochemical processes of the LOV1-domain of the blue-light receptor Phot1 from Chlamydomonas. We propose to analyze the electron transfer process that has been assumed to occur in the triplet state of LOV1 (FMN-type) and to study the subsequent formation of the signaling states in Phot1.A highly interdisciplinary approach combining molecular biology, biochemistry, spectroscopy, crystallography, and theory will be undertaken. Second, we plan to study sensory blue light receptors containing the so-called BLUF domain as chromophores (FAD-type). We will express the AppA-Protein, which is involved in the regulation of genes for photosynthetic proteins in the purple bacterium R.sphaeroides, and PAC-a/b that function as light-regulated adenylate cyclases in the flagellate E.gracilis. We will characterize the light activated enzymatic activities and try to determine the structures of both proteins in the dark form, and the light-activated state. We will study the primary photoprocesses by time and wavelength resolved UV-Vis and fluorescence spectroscopy, in particular in the ns time range. The spectral and kinetic information will be combined with the structural data to describe in detail the physical and photochemical reactions involved in the formation of the signaling states.