Mechanism of the staphylococcal pore-forming cytolytic toxins

葡萄球菌成孔溶细胞毒素的机制

• 批准号: 17380050
• 负责人: KAMIO Yoshiyuki
• 金额: $ 9.6万
• 依托单位: Environmental and Disaster prevention Research Institute, Tohoku Gakuin University (2006)Tohoku University (2005)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17380050/
• 关键词: staphylococcal toxin; two-component toxins; gamma-hemolysin; pore-forming; single molecule visualization; environment-sensitive fluoreophore; Recognition of target cells; Staphylococcus aureus; γヘモリジン; 2成分性溶血毒素; 膜孔形成機構; 標的細胞認識

Single-molecular imaging of the entrance of a protein into the hydrophobic environment of a cell membrane was investigated. The pre-stem of LukF, one of the two components of the pore-forming toxin staphylococcal gamma-hemolysin, was specifically labeled with 6-bromoacetyl-2-dimethylaminonaphthalene (Badan), an environment-sensitive fluorophore. Incubation of this derivative with erythrocyte an into the hydrohobic interior of the lipid bilayers. However, the increase in fluorescence was completely dependent on the interaction of Badan-labeled LukF with the gamma-hemolysin second component. Individual spots of Badan fluorescence on erythrocyte membrane were visualized that were associated with single pores. Analyses of the intensities of these fluorescent spots and their photobleaching independently showed that a single pore contained 3-4 LukF molecules. Thus, environment-sensitive fluorophore signals can be used to study the insertion of specific protein domains into cell membranes at the single-molecule level, and the use of this approach in the present study revealed that a single gamma-hemolysin pore opening contains at least three LukF molecules.

研究了蛋白质进入细胞膜疏水环境的单分子成像。 LukF 是成孔毒素葡萄球菌γ-溶血素的两种成分之一，其前茎被 6-溴乙酰基-2-二甲氨基萘 (Badan)（一种环境敏感荧光团）特异性标记。将该衍生物与红细胞一起孵育到脂质双层的疏水内部。然而，荧光的增加完全取决于 Badan 标记的 LukF 与 γ-溶血素第二组分的相互作用。红细胞膜上与单个孔相关的巴丹荧光的单个点被可视化。对这些荧光点的强度及其光漂白的独立分析表明，单个孔含有 3-4 个 LukF 分子。因此，环境敏感的荧光团信号可用于在单分子水平上研究特定蛋白质结构域插入细胞膜的情况，并且在本研究中使用这种方法表明，单个γ-溶血素孔开口至少包含三个 LukF 分子。

项目成果

Expression of g-hemolysin regulated by sae in Staphylococcus aureus strain Smith 5R

sae调控金黄色葡萄球菌Smith 5R中g-溶血素的表达

• 发表时间: 2006
• 期刊: FEMS Microbial.Lett (印刷中)
• 作者: Kazuko Yamazaki;Fuminori Kato;Yoshiyuki Kamio;Jun Kaneko
• 通讯作者: Jun Kaneko

Assembly of staphylococcal leukocidin into a pore-forming oligomer detergent-resistant membrane micro domains, lipid rafts, in human polymorphonuclear leukocytes

在人多形核白细胞中将葡萄球菌杀白细胞素组装成成孔低聚物、耐去垢剂膜微结构域（脂筏）

• 发表时间: 2006
• 期刊: Biosci. Biotechnol. Biochem. 70 (6)
• 作者: A.Nishiyama;J.Kaneko;M Harata;Y.Kamio
• 通讯作者: Y.Kamio