Structural Analysis of Intact Biomolecules by Solid State High Resolution Nuclear Magnetic Resonance with Homogenously Isotope Labeled Samples

通过固态高分辨率核磁共振对同位素标记样品的完整生物分子进行结构分析

• 批准号: 12450341
• 负责人: NOZAWA Tsunenori
• 金额: $ 9.41万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12450341/
• 关键词: Solid state high resolution NMR; Chlorosome; Bacteriochlorophyll c; Dimer; RFDR; Photosynthetic antenna; MCD; Isotope label; タイマー; 固体高分解能NMR; クロロゾーム

We have established the system for the structural analysis of intact biomolecules by solid state high resolution nuclear magnetic resonance (NMR) with homogenously isotope labeled samples. The system was applied to the membrane antenna LH1 of purple photosynthetic bacteria, and to the chlorosome antenna of green photosynthetic bacteria. The homogenously isotope labeled samples were obtained by cultivation of the bacteria with medium prepared with isotope enriched compounds. The membrane antennae LH1 was preliminary investigated by high resolution NMR in the solution dissolved in deutrated detergent, octyl-β-D-glucopyranoside. Many NMR signals were resolved and assigned for bacteriochlorophyll a due to the effects of the isotope enrichment. The model chlorosomes have been prepared with the isotope labeled bacteriochlorophyll c by the solvent vaporization method found by us. Application of the systems to the model chlorosomes revealed highly resolved signal, each of which appeared in pairwise. This implied that the chlorosomes are composed basic dimmer building block, or exist as two distinctly different conformations.

我们建立了一套利用固体高分辨核磁共振（NMR）分析完整生物分子结构的系统。将该系统应用于紫色光合细菌的膜天线LH 1和绿色光合细菌的叶绿体天线。通过用同位素富集化合物制备的培养基培养细菌获得均匀同位素标记的样品。用高分辨核磁共振技术对膜触角LH 1在含氘洗涤剂辛基-β-D-吡喃葡萄糖苷溶液中的结构进行了初步研究。由于同位素富集的影响，许多NMR信号被解析并分配给细菌叶绿素a。用同位素标记的细菌叶绿素c，用我们发现的溶剂蒸发法制备了模型绿质体。应用该系统的模型叶绿体显示高分辨率的信号，其中每一个成对出现。这意味着叶绿体是由基本的二聚体结构单元组成的，或者是以两种截然不同的构象存在。