Molecular study on large conformational changes of protein that relates biofunction and diseases

与生物功能和疾病相关的蛋白质大构象变化的分子研究

• 批准号: 15370047
• 负责人: KAWATA Yasushi
• 金额: $ 8.32万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15370047/
• 关键词: Amyloid fibril; Oligomeric protein; α-synuclein; Molecular chaperone; Molecular compactness; Functional mechanism; Chaperonin; Neurodegenerative disease; GroES; コンフォメーション病; 構造変化; 構造安定性; 蛋白質の構造安定性; コンフォメーション変化; アミロイド線維形成; シャペロニンGroES

Studies on protein stability including global conformational changes and functional mechanism of molecular chaperone, which are closely related to biofunction and diseases, were performed in detail and following results were obtained.1.Study on structure and stability of oligomeric protein : We have determined X-ray crystal structure of thermostable aspartase enzyme, and elucidated the mechanism of thermostability and active site structure of the enzyme comprising from 4 identical subunits. On the other hand, we studied solution structure and molecular unfolding mechanism of E.coli co-chaperonin GroES heptamer at high protein concentrations by using small angle X-ray scattering.2.Study on conformational changes and amyloid fibril formation of protein : We have found that oligomeric protein GroES, that is a non-related protein to disease, formed a typical amyloid fibril under a certain condition, and elucidated the fibril formation mechanism in terms of molecular compactness. Furthermore, we studied fibril formation mechanism of α-synuclein, a causative protein of Parkinson disease, and proved that the amyloid fibril formation of α-synuclein is accelerated markedly in the presence of preformed seeds of other different protein's fibrils.3.Study on structure and function of molecular chaperone : We have studied in detail structure and function relationship of group I chaperonin GroEL from E.coli and group II chaperonins from hyper-thermostable strains. We have found that domain movements of GroEL are very important for the function and that cobalt and manganese ions are novel factors for nucleotide hydrolysis activity and substrate refolding function of group II chaperonin. Furthermore, we have investigated function of zebrafish Hsp60, and elucidated that Hsp60 is required for blastema formation and maintenance during regeneration.

对与生物功能和疾病密切相关的蛋白质稳定性，包括分子伴侣的整体构象变化和功能机制进行了详细的研究，得到了以下结果。 1.寡聚蛋白的结构和稳定性研究：我们测定了耐热天冬氨酸酶的X射线晶体结构，阐明了该酶的热稳定性机制和活性位点结构 4个相同的亚基。另一方面，我们利用小角X射线散射研究了大肠杆菌共伴侣蛋白GroES七聚体在高蛋白质浓度下的溶液结构和分子解折叠机制。2.蛋白质构象变化和淀粉样原纤维形成的研究：我们发现寡聚蛋白GroES是一种与疾病无关的蛋白质，在一定浓度下形成了典型的淀粉样原纤维。 条件，并阐明了分子致密性方面的原纤维形成机制。此外，我们还研究了帕金森病致病蛋白α-突触核蛋白的原纤维形成机制，并证明在其他不同蛋白原纤维预制种子的存在下，α-突触核蛋白的淀粉样原纤维形成显着加速。3.分子伴侣的结构和功能研究：我们详细研究了I组的结构和功能关系。 来自大肠杆菌的伴侣蛋白 GroEL 和来自超热稳定菌株的 II 组伴侣蛋白。我们发现 GroEL 的结构域移动对其功能非常重要，并且钴和锰离子是 II 族伴侣蛋白的核苷酸水解活性和底物重折叠功能的新因子。此外，我们还研究了斑马鱼Hsp60的功能，并阐明了Hsp60是再生过程中胚基形成和维持所必需的。

项目成果

T.Fujii et al.: "Crystal Structure of Thermostable Aspartase from Bacillus sp. YM55-1: Structure-based Exploration of Functional Sites in the Aspartase Family"Journal of Molecular Biology. 328・3. 635-654 (2003)

T.Fujii 等：“来自芽孢杆菌 YM55-1 的耐热天冬氨酸酶的晶体结构：基于结构的天冬氨酸酶家族功能位点探索”《分子生物学杂志》328・3（2003 年）。

A Novel ATP/ADP Hydrolysis Activity of Hyperthermostable Group II Chaperonin in the Presence of Cobalt or Manganese Ion

钴或锰离子存在下超热稳定 II 族伴侣蛋白的新型 ATP/ADP 水解活性

• 发表时间: 2006
• 期刊: FEBS Letters 580
• 作者: 遠藤斗志也;吉久 徹;森 和俊;田口英樹;Kunihiro Hongo et al.
• 通讯作者: Kunihiro Hongo et al.

Induction of AApoAII amyloidosis by various heterogenous amyloid fibrils

各种异质淀粉样原纤维诱导 AApoAII 淀粉样变性

• 发表时间: 2004
• 期刊: FEBS Letters 563
• 作者: X.Fu et al.

Amyloid Fibril Formation of a-Synuclein is Accelerated by Preformed Amyloid Seeds of Other Proteins : Implications for the Mechanism of Transmissible Conformational Diseases

其他蛋白质的预制淀粉样蛋白种子加速了α-突触核蛋白的淀粉样原纤维形成：对传染性构象疾病机制的影响

• 发表时间: 2005
• 期刊: J. Biol. Chem. 280・46
• 作者: Hisashi Yagi;Eiko Kusaka;Kunihiro Hongo;Tomohiro Mizobana;Yasushi Kawata
• 通讯作者: Yasushi Kawata

Hsp60 is Required for Blastema Formation and Maintenance during Regeneration

Hsp60 是再生过程中胚基形成和维持所必需的

• 发表时间: 2005
• 期刊: Proc. Natl. Acad. Sci. USA 102・41
• 作者: Shinji Makino;Geoffrey G.Whitehead;Ching-Ling Lien;Akane Kono;Yasushi Kawata;Mark T.Keating
• 通讯作者: Mark T.Keating