Development of New Methodologies for Protein-Interaction Analysis

蛋白质相互作用分析新方法的开发

• 批准号: 10179101
• 负责人: AKUTSU Hideo
• 金额: $ 84.61万
• 依托单位: Osaka University (2000-2001)Yokohama National University (1998-1999)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas (A)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10179101/
• 关键词: Protein interaction; Protein segmental labeling; NMR; Solution structure determination; Magic angle sample spinning; Solid state NMR; Ab initio X-ray crystallography; Thioester method; 部分セグメント標識法; 立体・位置特異的重水素化; 固体高分解能多次元NMR; 安定同位体標識; X線溶液散乱法 /

1) A new segmental isotope labeling method has been developed for structural analysis of large proteins. This method was successfully applied to RNA polymerase α subunit C-domain, maltose-binding protein, ATPase β-subunit.2) A series of new techniques in solid state NMR have been developed for structural analysis of large proteins and membrane proteins that are unsuitable for the analysis by X-ray crystallography and solution NMR.3) An Ab initio method for X-ray crystal analysis have been developed for analysis of huge biological systems.4) A thioester method has been developed for chemical synthesis of proteins to investigate protein-protein interactions.

1）已开发了一种新的分段同位素标记方法，用于大蛋白的结构分析。该方法成功地应用于RNA聚合酶α亚基C-蛋白酶，麦芽糖结合蛋白，ATPaseβ-亚基。2）2）一系列固态NMR中的一系列新技术已被用于大型蛋白质和膜蛋白的结构分析，用于对X型 - ray Crybay的分析不适合进行X-RAY CRYER.3 INSER.3的分析。3巨大的生物系统。4）已经开发了一种用于蛋白质化学合成以研究蛋白质 - 蛋白质相互作用的方法。

项目成果

K.Haneda,: "Chemo-enzymatic synthesis of calcitonin derivatives containing N-linked origosaccharides" Bioorg.Medi.Chem.Lett.8. 1303-1306 (1998)

K.Haneda，：“含有N-连接低聚糖的降钙素衍生物的化学酶合成”Bioorg.Medi.Chem.Lett.8。

K. Nomura: "Determination of the Complete Structure of a Uniformly Labeled Molecule by Rotational Resonance Solid-State NMR in the Tilted Rotational Frame"J. Am. Chem. Soc.. 121. 4064-4065 (1999)

K. Nomura：“通过倾斜旋转框架中的旋转共振固态核磁共振测定均匀标记分子的完整结构”J。

K.Tozawa 外5名: "Functions and ATP-binding responses of the twelve histidine residues in the TF_1-ATPase β subunit"J. Biochem.. 130. 527-533 (2001)

K. Tozawa 和其他 5 人：“TF_1-ATPase β 亚基中十二个组氨酸残基的功能和 ATP 结合反应”J. Biochem.. 130. 527-533 (2001)

R.Riek 外4名: "[^<13>C,^<13>C] -and [^<13>C,^1H] -TROSY in a Triple Resonance Experiment for Ribose -base Intrabase Correlations in Nuclei Acids"J. Am. Chem. Soc.. 123. 658-664 (2001)

R.Riek 和其他 4 人：“[^<13>C,^<13>C] - 和 [^<13>C,^1H] -TROSY in a Triple Resonance Experiment for Ribose-base Intrabase Correlations in Nucleic Acids”化学杂志。123。658-664（2001）

Ida, K.: "The 1.55 A resolution structure of Nicotiana alata S_<F11>-Rnase associated with gametophytic self-incompatibility"J. Mol. Biol.. Vol. 314. 103-112 (2001)

Ida, K.：“与配子体自交不亲和性相关的烟草 S_<F11>-Rnase 的 1.55 A 解析结构”J。