Structural basis of the interplay between the carbohydrate recognition and protein ubiquitination systems in the glycoprotein degradation in cells

细胞糖蛋白降解中碳水化合物识别和蛋白质泛素化系统之间相互作用的结构基础

• 批准号: 18390016
• 负责人: KATO Koichi
• 金额: $ 11.21万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390016/
• 关键词: Protein degradation; Glycan; Ubiquitination; NMR; Structural biology

For a better understanding the molecular basis for the underlying mechanisms of the intracellular glycoprotein degradation governed by glycosylation and ubiquitination, we used a fusional approach integrating the methodologies of glycobiology and structural biology.1. Elucidation of sugar-binding properties of the intracellular lectins by using a sugar library.We constructed a high-mannose-type sugar library using the multi-dimensional HPLC method and applied them to frontal affinity chromatography (FAC) analyses of sugar-binding specificities of a variety of intracellular lectins involved in the quality control of glycoproteins in cells. The FAC data revealed that the sugar-binding domains of the endoplasmic reticulum (ER) chaperones, cargo receptors, ubiquitin ligases, and deglycosylation enzyme specifically recognize distinct glycotopes displayed on the partially trimmed high-mannose-type oligosaccharides in the N-glycan processing pathway and thereby determine the glycoprotein-fate … More s in cells.2. Elucidation of the molecular mechanisms of the glycoprotein degradation system by structural biology approaches. On the basis of the stable-isotope-assisted NMR data, we elucidated the structural basis of the molecular recognition by the sugar-binding domain of the ubiquitin ligase SCFFbs1, which ubiquitinates malfolded glycoproteins retrogradely transferred from the ER to the cytosol, and by the ubiquitin-binding domain of the deglycosylation enzyme peptide:Nglycanase, which facilitates the proteasomal degradation of glycoproteins. Furthermore, we determined the ubiquitin recognition modes of ubiquitin ligases and de-ubiquitinating enzymes by using a library of ubiquitin chains. In addition, our NMR data revealed the mechanisms of the activation of the ubiquitin-ligase complexes upon covalent attachment of Nedd8. These results provide insights into the molecular basis of the interplay between the carbohydrate recognition and protein ubiquitination systems in the glycoprotein degradation in cells. Less

为了更好地理解由糖基化和泛素化控制的细胞内糖蛋白降解的基本机制，我们使用了整合糖基生物学和结构生物学方法的融合方法。1。通过使用糖内库来阐明细胞内讲座的糖结合特性。我们使用多维HPLC方法构建了高臭型糖库，并将其应用于额叶亲和色谱法（FAC）分析，用于各种细胞内质量控制的细胞内糖内讲座的糖结合规范。 FAC数据表明，内质网（ER）伴侣的糖结合域，货物接收器，泛素连接酶和去糖基化酶在部分修剪的高含量寡糖含量上，在N-Glyculy中均具有较高的含量，并在其中识别了n-glyccus，并在其中识别了一个杂物，并在其中识别了杂物，并在其中识别出了良好的含量……单元格2。通过结构生物学方法阐明糖蛋白降解系统的分子机制。 On the basis of the stable-isotope-assisted NMR data, we elucidated the structural basis of the molecular recognition by the sugar-binding domain of the ubiquitin ligase SCFFbs1, which ubiquitinates malfolded glycoproteins retrogradely transferred from the ER to the cytosol, and by the ubiquitin-binding domain of the deglycosylation enzyme胡椒：nglycanase，促进糖蛋白的蛋白酶体降解。此外，我们通过使用泛素链库来确定泛素连接酶和去泛素化酶的泛素识别模式。此外，我们的NMR数据揭示了NEDD8共同附着后，泛素 - 连接酶复合物激活的机制。这些结果为细胞中糖蛋白降解中碳水化合物识别和蛋白质泛素化系统之间相互作用的分子基础提供了见解。较少的

项目成果

NMR structural biology of protein quality control in cells

细胞内蛋白质质量控​​制的核磁共振结构生物学

• 发表时间: 2007
• 作者: Kurose;H;Koichi Kato
• 通讯作者: Koichi Kato

An NMR approach to the protein society in cells

细胞中蛋白质社会的核磁共振方法

• 发表时间: 2006
• 作者: Koichi;Kato
• 通讯作者: Kato

Crystal structure of a chaperone complex that contributes to the assembly of yeast 20S proteasomes

• DOI: 10.1038/nsmb.1386
• 发表时间: 2008-03-01
• 期刊: NATURE STRUCTURAL & MOLECULAR BIOLOGY
• 影响因子: 16.8
• 作者: Yashiroda, Hideki;Mizushima, Tsunehiro;Tanaka, Keiji
• 通讯作者: Tanaka, Keiji

Comprehensive Glycoscience

综合糖科学

• 发表时间: 2007
• 作者: Oohashi T;Bekku Y;Houliston RS;Van Sorge NM;Van Sorge NM;Overell J;Koga M;Kamitani T;Tatsumoto M;Nagasawa K;Houliston RS;Yoshida T;Funakoshi K;Tatsumoto M;Komagamine T;Gono T;Yuki N;Kimoto K;Comin R;Koga M;Koga M;Koga M;Pandey JP;Yuki N;Li J;Yuki N;Odaka M;Odaka M;Koga M;Hiraga A;Kuwabara S;Goodfellow JA;Takahashi M;Tatsumoto M;Kamitani T;S. Matsumiya;K. Kato;S. Matsumiya;K. Kato;Y.Wada;矢木宏和;S.Matsumiya;山口芳樹;C.-T.Guo;K.Kato;M.Holland;Y.Yamaguchi;K.Talabnin;Y.Kamiya;S.-J.Yoon;Y.Yamaguchi et al.;M.Holland et al.;Y.Yamaguchi et al.;Y. Kamiya;Y. Kamiya;T.Kimura et al.;H.Yagi et al.;Y.Kamiya et al.;S.Sekiya et al.;N.Takahashi
• 通讯作者: N.Takahashi

細胞内タンパク質社会の理解を目指したNMR構造生物学

核磁共振结构生物学旨在了解细胞内蛋白质社会

• 发表时间: 2006
• 作者: Koichi;Kato;加藤 晃一
• 通讯作者: 加藤 晃一