Structural and functional analysis of a cell wall-associated giant protein EbhA from Staphylococcus aureus

金黄色葡萄球菌细胞壁相关巨蛋白 EbhA 的结构和功能分析

• 批准号: 18310141
• 负责人: TANAKA Isao
• 金额: $ 11.56万
• 依托单位: Hokkaido 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-18310141/
• 关键词: Staphylococcus aureus; Cell-wall associated protein; Adhesion mechanism; X-ray structure analysis; Binding protein

This research focused on the protein involved in the adhesion of Staphylococcus aureus to the host cell. The final goal of this research is to understand the adhesion mechanism of this bacterium to the host cell and to establish the structural basis for the drug design. During the project period, the structure of a giant protein EbhA, which was thought to be involved in the endocarditis, was investigated by the X-ray crystallography, small angle X-ray scattering, CD spectroscopy and electron microscopy. First, we elucidated the structure of the two repeating units (Ebh-R7-R8) of this molecule by the X-ray crystallography. Then, based on the studies of the small angle X-ray scattering and CD spectrometry, we proposed that these structures are connected tandem. Furthermore, the electron microscopy showed that the Ebh-R10-19 is the string of about 500A in length which is consistent with the proposed structure. Finally, we simulated the whole Ebh structure by calculating the helical parameters of this molecule based on the X-ray structure, which showed that Ebh is the string of about 3200 A in length. These studies suggest that Ebh is on the surface of the Staphylococcus aureus cell and plays the role as "skeleton molecule" to reinforce the cell.

这项研究的重点是参与金黄色葡萄球菌与宿主细胞粘附的蛋白质。这项研究的最终目标是了解这种细菌与宿主细胞的粘附机制，并为药物设计奠定结构基础。项目期间，通过X射线晶体学、小角X射线散射、CD光谱和电子显微镜研究了被认为与心内膜炎有关的巨型蛋白EbhA的结构。首先，我们通过X射线晶体学阐明了该分子的两个重复单元（Ebh-R7-R8）的结构。然后，基于小角X射线散射和CD光谱的研究，我们提出这些结构是串联的。此外，电子显微镜显示Ebh-R10-19是长度约为500A的线，这与所提出的结构一致。最后，我们根据X射线结构计算了该分子的螺旋参数，模拟了整个Ebh结构，结果表明Ebh是长度约为3200 A的弦。这些研究表明，Ebh位于金黄色葡萄球菌细胞的表面，发挥着“骨架分子”的作用，增强了细胞的强度。

项目成果

巨大細胞表面蛋白質Ebhの全体構造に関する考察

巨细胞表面蛋白Ebh整体结构探讨

• 发表时间: 2008
• 作者: 田中良和

A Helical String of Alternately Connected Two Three-helix Bundles for the 1.1-Megadalton Cell Wall-Associated Adhesion Protein Ebh from Staphylococcus aureus

来自金黄色葡萄球菌的 1.1 兆道尔顿细胞壁相关粘附蛋白 Ebh 的一串交替连接的两个三螺旋束

• 发表时间: 2008
• 作者: Yoshikazu Tanaka

Structural analysis of a giant cell wall-associated protein Ebh from Staphylococcus aureus

金黄色葡萄球菌巨细胞壁相关蛋白 Ebh 的结构分析

• 发表时间: 2008
• 作者: Yoshikazu Tanaka

A Helical String of Alternately Connected Two Threehelix Bundles for the 1.1-Megadalton Cell Wall-Associated Adhesion Protein Ebh from Staphylococcus aureus

来自金黄色葡萄球菌的 1.1 兆道尔顿细胞壁相关粘附蛋白 Ebh 的交替连接的两个三螺旋束的螺旋串

• 发表时间: 2008
• 作者: Yoshikazu Tanaka

ブドウ球菌表層に存在する1.1-Megadalton細胞接着蛋白質Ebhの構造解析

葡萄球菌表面 1.1-Megadalton 细胞粘附蛋白 Ebh 的结构分析

• 发表时间: 2008
• 作者: 田中良和