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

蓝光光感受器的时间分辨光谱表征

• 批准号: 5420251
• 负责人: Professor Dr. Bernhard Dick
• 金额: --
• 依托单位: Institut für Experimentelle und Angewandte Physik
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2004
• 资助国家: 德国
• 起止时间: 2003-12-31 至 2010-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5420251?language=en
• 关键词: Time; resolved; spectroscopic; characterization; blue

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.

我们已经分析了衣原体蓝光受体Phot1的Lov1域的结构和光化学过程。我们建议分析已假定在LOV1（FMN类型）三重态的电子传递过程，并研究PHOT1中信号态的随后形成。将进行分子生物学，生物化学，光谱，光谱，晶体学和理论的高度跨学科方法。其次，我们计划研究包含所谓的BLUF结构域作为发色团（FAD型）的感觉蓝光受体。我们将表达APPA蛋白，该蛋白参与了紫色细菌r.sphaeroides中光合蛋白的调节，而PAC-A/B则起作用，可作为鞭毛蛋白酶中光调节的腺苷酸环化酶起作用。我们将表征光激活的酶活性，并尝试以黑暗形式和光激活状态确定两种蛋白质的结构。我们将按时间和波长分辨的UV-VIS和荧光光谱研究，特别是在NS时间范围内研究主要的光程序。光谱和动力学信息将与结构数据相结合，以详细描述信号态形成所涉及的物理和光化学反应。

项目成果

Blue‐Light Induced Interaction of LOV Domains from Chlamydomonas reinhardtii

蓝光诱导莱茵衣藻 LOV 结构域的相互作用

• DOI: 10.1002/cbic.200800158
• 发表时间: 1938
• 期刊: ChemBioChem
• 影响因子: 3.2
• 作者: R. J. Kutta;E. S. A. Hofinger;H. Preuss;G. Bernhardt;B. Dick
• 通讯作者: B. Dick