Molecular basic research on protein aggregation and conformational diseases

蛋白质聚集与构象疾病的分子基础研究

基本信息

  • 批准号:
    12680613
  • 负责人:
  • 金额:
    $ 2.24万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
  • 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

项目摘要

Protein aggregation has become one of the serious problems in the fields of biotechnology, medicine, and life science. In this study, we have studied basic mechanism of insoluble protein aggregation in vivo and in vitro by using various protein molecular-scientific techniques.1. Characterization of folding intermediate of triose phosphate isomerase(TIM)It has been elucidated that the folding intermediate of dimeric enzyme TIM was a partially structured monomeric species by NMR spectroscopic analysis.2. Solubilization of inclusion body of thermostable Mn-catalaseInsoluble aggregations (inclusion body) were formed when Mn-catalase was overexpressed in E. coli. However, it has been elucidated that co-expression of molecular chaperones (GroEL, GroES, DnaK, DnaJ, and GrpE) overcome the situation, and the correct folded active enzyme was obtained efficiently.3. Structural changes and amyloid-like fibril formation of GroES and α synucleinIncubated chaperonin GroES, a heptameric protein, at a high protein concentration for several days in the presence of Gdn-HCl, global conformational changes occurred and formed insoluble amyloid-like fibrils, which has been elucidated by Thioflavin-T staining and the electron microscopy. In addition, it was observed that compaction of natively unfolded α synuclein by increasing ionic strength led to amyloid fibril formation readily. These results gave a significant insight on mechanism of conformational changes of various proteins related to insoluble aggregations and amyloid-like fibril formation.
蛋白质聚集已成为生物技术、医学和生命科学领域的严重问题之一。在本研究中,我们利用多种蛋白质分子科学技术,研究了不溶性蛋白质在体内和体外聚集的基本机制。磷酸三糖异构酶(TIM)折叠中间体的表征通过核磁共振波谱分析,证明二聚体酶TIM的折叠中间体为部分结构单体物种。耐热锰过氧化氢酶包涵体的增溶作用当锰过氧化氢酶在大肠杆菌中过表达时,形成难溶的聚集体(包涵体)。然而,已阐明分子伴侣(GroEL、GroES、Dna K、Dna J和GRPE)的共表达克服了这种情况,并有效地获得了正确的折叠活性酶。Groes和α突触核团的结构变化和淀粉样原纤维的形成分子伴侣蛋白是一种七聚体蛋白,在GdN-HCl存在下,在高蛋白浓度下孵育几天后,全局构象发生变化,形成不溶性的淀粉样原纤维,硫代黄素-T染色和电子显微镜证实了这一点。此外,还观察到自然展开的α突触核蛋白通过增加离子强度而压实,容易导致淀粉样原纤维的形成。这些结果对与不溶性聚集和淀粉样原纤维形成相关的各种蛋白质的构象变化机制提供了重要的见解。

项目成果

期刊论文数量(6)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Mizobata,T. et al.: "Cloning and Over-expression of Thermostable Bacillus sp. YM55-1 Aspartase and Site-directed Mutagensis for Probing a Catalytic Residue"Eur.J.Biochem.. 267・6. 1847-1857 (2000)
Mizobata, T. 等:“用于探测催化残基的耐热芽孢杆菌属 YM55-1 天冬氨酸酶的克隆和过表达以及定点诱变”Eur.J.Biochem.. 267・6(2000 年) )
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Morgan,C.J. et al.: "A Compact Monomeric Intermediate Identified by NMR in the Denaturation of Dimeric Triose Phosphate Isomerase"J.Mol.Biol.. 300・1. 11-16 (2000)
Morgan, C.J. 等:“二聚丙糖磷酸异构酶变性中通过 NMR 鉴定的紧凑单体中间体”J.Mol.Biol.. 300・1 (2000)。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Mizobata, T. et al.: "Overproduction of Thermus sp. Ys 8-13 Manganese Catalase in Escherichia coli : Production of Soluble Apo Enzyme and In Vitro Formation of Active Holo Enzyme"European Journal of Biochemistry. 267・13. 4264-4271 (2000)
Mizobata, T. 等人:“大肠杆菌中栖热菌 Ys 8-13 锰过氧化氢酶的过量生产:可溶性 Apo 酶的生产和活性全息酶的体外形成”《欧洲生物化学杂志》267·13。 (2000)
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Morgan, C.J. et al.: "A Compact Monomeric Intermediate Identified by NMR in the Denaturation of Dimeric Triose Phosphate Isomerase"Journal of Molecular Biology. 300・1. 11-16 (2000)
Morgan, C.J. 等人:“二聚丙糖磷酸异构酶变性中通过 NMR 鉴定的紧凑单体中间体”,分子生物学杂志 300・1 (2000)。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

KAWATA Yasushi其他文献

KAWATA Yasushi的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('KAWATA Yasushi', 18)}}的其他基金

Structural and functional characteristics of natively unfolded protein
天然未折叠蛋白质的结构和功能特征
  • 批准号:
    21570113
  • 财政年份:
    2009
  • 资助金额:
    $ 2.24万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
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
  • 财政年份:
    2003
  • 资助金额:
    $ 2.24万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Protein conformational changes and molecular chaperone
蛋白质构象变化和分子伴侣
  • 批准号:
    14037241
  • 财政年份:
    2002
  • 资助金额:
    $ 2.24万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了