Structural investigation of aggregates in various functional complexes, polymers and biomolecules hi solution by using PFG-NMR analysis.

使用 PFG-NMR 分析对溶液中各种功能复合物、聚合物和生物分子中的聚集体进行结构研究。

• 批准号: 14571995
• 负责人: SEKI Hiroko
• 金额: $ 1.15万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571995/
• 关键词: PFG NMR; cold spray ionization (CSI)-MS; diffusion; hydrogen bonding; chain structure; steroid compounds; X-ray analysis; protein-ligand complex; HMBC; 分子クラスター; 生体分子; 緩和時間; パルスシーケン

PFG NMR, especially used for diffusion experiment, and CSI (cold-spray ionization)-MS has been our major subject for solution structure analysis in recent years. X-ray analysis elucidating solid structure is also contributed to this study in some cases. It was found that some steroid compounds behave as the large-scale aggregated chain structure in solution. This unprecedented phenomenon was definitely confirmed by using CSI-MS and PFG NMR Characteristic chain structures based on strong intermolecular hydrogen bonding in the case of cortisone was clearly observed in solution exhibiting series of plural clusters, which was assigned up to thirteen aggregates. On the other hand, progesterone slightly weak intermolecular interaction was predicted form CSI-MS because ion peaks assigned only up to trimeric cluster. This result is identical with that obtained from X-ray analysis. Recently, it was demonstrated that NMR diffusion studies is an useful method for the characterization of a variety … More of interaction system in solution. NMR diffusion experiments were carried out on a LA-600 by using BPP-STE method. Calibration curve obtained from observed diffusion coefficients and calculated molecular volume. In the case of progesterone, observed diffusion coefficient (6.98×10^<-10>m^2/s) is as nearly same as calculated. The reason for this result indicated that progesterone has no intermolecular interaction in solution as well as in the crystal. On the contrary, diffusion coefficient of cortisone (5.58×10^<-10>m^2/s) exhibits slightly different from calculated values, presumably because hydrogen bonding occurs to afford molecular cluster. In consequence, these steroid compounds appeared to maintain ordered clusters, based on hydrogen, bonding in diluted solution, instead diffused randomly as individual molecules, as has been generally accepted.Multidimensional T_<1ρ^->, diffusion-filtered and diffusion-ordered NOESY techniques are very suitable techniques for identifying segments of a ligand binding with a protein receptor in the drug discovery process. Less

PFG NMR（尤其是用于扩散实验的PFG NMR）和CSI（冷喷雾电离）-MS是近年来我们在溶液结构分析方面的主要研究课题。在某些情况下，阐明固体结构的X射线分析也有助于这项研究。研究发现，某些甾体化合物在溶液中表现为大尺度聚集的链状结构。通过使用CSI-MS和PFG NMR明确证实了这种前所未有的现象。在可的松的情况下，在溶液中清楚地观察到基于强分子间氢键的特征链结构，其表现出一系列的复数簇，其被分配为多达13个聚集体。另一方面，从CSI-MS预测孕酮的分子间相互作用略弱，因为离子峰仅归属于三聚体簇。这一结果与X射线分析结果一致。最近，它被证明，NMR扩散研究是一种有用的方法，用于表征各种 ...更多信息 相互作用系统的最大值。用BPP-STE方法在LA-600上进行了NMR扩散实验。从观察到的扩散系数和计算的分子体积获得的校准曲线。在孕酮的情况下，观察到的扩散系数（6.98×10^<-10>m^2/s）与计算值几乎相同。这一结果的原因表明，孕酮在溶液中以及在晶体中没有分子间相互作用。与此相反，可的松的扩散系数（5.58×10^<-10>m^2/s）与计算值略有不同，可能是由于氢键作用形成了分子簇。结果表明，这些甾体化合物在稀溶液中以氢键为基础形成有序的簇，而不是像人们普遍认为的那样以单个分子的形式随机扩散.多维T_&lt;1ρ^-&gt;、扩散过滤和扩散有序NOESY技术是在药物发现过程中识别配体与蛋白质受体结合片段的非常合适的技术.少

项目成果

H.Utsumi, H.Seki, K.Yamaguchi, M.Tashiro: "Segment Identification of a Ligand binding with a Protein Receptor Using Multidimentional T1 ρ -, Diffusion-Filtered, and Diffusion-Ordered NOESY Experimentas"Anal.Sci.. 19. 1441-1443 (2003)

H.Utsumi、H.Seki、K.Yamaguchi、M.Tashiro：“使用多维 T1 ρ -、扩散过滤和扩散有序 NOESY 实验对与蛋白质受体结合的配体进行片段鉴定”Anal.Sci.. 19 .1441-1443 (2003)

Y.Sei: "Observation of Waters Bound to a Protein Using Temperature Dependent Cold-Spray Ionization Mass Spectrometry"Angew.Chem.Int.Ed., revised Manuscript (No. Z 53918). (2004)

Y.Sei：“使用温度依赖性冷喷雾电离质谱法观察与蛋白质结合的水”Angew.Chem.Int.Ed.，修订手稿（编号 Z 53918）。

K.Shikii, S.Sakamoto, H.Seki, H.Utsunii, K.Yamaguchi: "Narcissistic aggregation of steroid compounds in diluted solution elucidated by CSI-MS, PFG NMR and X-ray analysis"Tetrahedron. 60. 3487-3492 (2004)

K.Shikii、S.Sakamoto、H.Seki、H.Utsunii、K.Yamaguchi：“通过 CSI-MS、PFG NMR 和 X 射线分析阐明稀释溶液中类固醇化合物的自恋聚集”四面体。

M.-J Cheng, B.Jayaprakasam, T.Ishikawa, H.Seki, I.-L.Tsai, J.-J.Wang, I.-S Chen: "Chemical and Cytotoxic Constituents from the Stem of Machilus zuihoensis"Helv.Chim.Acta.. 85. 1909-1914 (2002)

M.-J Cheng、B.Jayaprakasam、T.Ishikawa、H.Seki、I.-L.Tsai、J.-J.Wang、I.-S Chen：“瑞丰润楠茎的化学和细胞毒性成分”

H.Ttsumi, H.Seki, K.Yamaguchi, M.Tashiro: "Segment Identification of a Ligand binding with a Protein Receptor Using Multidimentional T_<1□>-, Diffusion-Filtered and Difufusion-OrderedNOESY Experimentas"Anal.Sci.. 19. 1441-1443 (2003)

H.Ttsumi、H.Seki、K.Yamaguchi、M.Tashiro：“使用多维 T_<1□>-、扩散过滤和扩散有序 NOESY 实验对与蛋白质受体结合的配体进行片段鉴定”Anal.Sci.. 19. 1441-1443 (2003)