MODE OF ACTION OF STAPHYLOCOCCAL LEUKOCIDIN AND gamma-HEMOLYSIN

金黄色葡萄球菌杀白细胞素和γ-溶血素的作用方式

• 批准号: 09670275
• 负责人: TOMITA Toshio
• 金额: $ 2.05万
• 依托单位: FACULTY OF AGRICULTURE,TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670275/
• 关键词: Staphylococcus aureus; bi-component cytolysin; staphylococcal gamma-hemolysin; staphylococcal leukocidin; pore-forming toxin; complex formation; ring structure; erythrocyte; neutrophil; リング状構造

Staphylococcal gamma-hemolysin consists of Hlg1 (or Luk F) and Hlg2, which cooperatively lyze human erythrocytes. Since gamma-hemolysin caused swelling of human erythrocytes before lysis, we studied pore-forming nature of the toxin by use of polyethylene glycols as osmotic protectants and determined the functional diameter of the pore. To elucidate the molecular architecture of the membrane pore formed by gamma-hemolysin, we solubilized the pore complex with 2 % sodium dodecyl sulfate, purified it by sucrose gradient centrifugation, and observed the purified pore complex under an electron microscope. Our data showed that the Hlg1 and the Hlg2 components assemble into a ring-shaped 200 kDa complex in a molar ratio of 1 : 1, forming a membrane pore with a functional diameter of 2.1-2.4 nm.Staphylococcal leukocidin consists of LukS and LukF, which lyse human and rabbit polymorphonuclear leukocytes and rabbit erythrocytes. Here we studied the pore-forming properties of leukocidin and the molecular architecture of the leukocidin pore. (1) Leukocidin caused an efflux ofpotassium ions from rabbit erythrocytes and swelling of the cells before lysis. However, ultimate lysis of the swollen cells did not occur when polyethylene glycols with hydrodynamic diameters of <greater than or equal> 2.1 nm were present in the extracellular space. (2) Electron microscopy showed that the leukocidin formed a ring-shaped structure with outer and inner diameters of 9 and 3 nm, respectively, on human polymorphonuclear leukocytes and rabbit erythrocytes. (3) Ring-shaped structures of the same dimensions were isolated from the target cells, and they contained LukS and LukF in a molar ratio of 1 : 1. (4) A single ring-shaped toxin complex had a molecular size of 205 kDa. These results indicated that LukS and LukF assemble into a ring-shaped oligomer of approximately 200 kDa on the target cells, forming a membrane pore with a functional diameter of approximat

葡萄球菌γ-羟糖蛋白蛋白由HLG1（或Luk F）和HLG2组成，它们合作地lyze人类红细胞。由于γ-脱糖蛋白在裂解前引起人类红细胞的肿胀，因此我们通过将聚乙烯乙二醇作为渗透保护剂研究了毒素的孔形成性质，并确定了孔的功能直径。为了阐明由γ-毛糖蛋白形成的膜孔的分子结构，我们用2％十二烷基硫酸钠溶解了孔复合物，通过蔗糖梯度离心纯化，并在电子显微镜下观察到纯化的孔复合物。 Our data showed that the Hlg1 and the Hlg2 components assemble into a ring-shaped 200 kDa complex in a molar ratio of 1 : 1, forming a membrane pore with a functional diameter of 2.1-2.4 nm.Staphylococcal leukocidin consists of LukS and LukF,裂解人类和兔子多形核白细胞和兔红细胞。在这里，我们研究了白细胞素的孔形成特性和白细胞蛋白孔的分子结构。 （1）白细胞素在裂解前引起了兔红细胞和细胞肿胀的腹腔离子的排出。但是，当细胞外空间中存在流体动力直径<大于或等于> 2.1 nm的聚乙烯乙二醇时，不会发生肿胀细胞的最终裂解。 （2）电子显微镜表明，白细胞素在人类多形核白细胞和兔红细胞上形成了一个环形结构，外径分别为9和3 nm。 （3）从目标细胞中分离出相同维度的环形结构，它们的摩尔比为1：1。（4）单个环形毒素复合物的分子大小为205 kDa 。这些结果表明，Luks和Lukf在目标细胞上组装成大约200 kDa的环形寡聚物，形成一个膜孔，具有大约功能直径的膜孔

项目成果

Sugawara,N., Tomita,T., and Kamio,Y.: "Assembly of Staphylococcus aureus γ-hamolysin into a pore-forming,ring-shapged complex on the surface of human erythrocytes" FEBS Lett.410. 333-337 (1997)

Sukawara, N.、Tomita, T. 和 Kamio, Y.：“金黄色葡萄球菌 γ-溶血素在人红细胞表面组装成成孔环形复合物” FEBS Lett.410（ 1997）

Sugawara, N., Tomita, T., Sato, T., and kamio, Y.: "Assembly of staphylococcal leukocidin into a pore-forming ring-shaped complex on rabbit erythrocytes and human polymorphonuclear leukocytes" Biosci.Biotechnol.Biochem.63・5印刷中. (1999)

Sukawara, N.、Tomita, T.、Sato, T. 和 kamio, Y.：“将葡萄球菌杀白细胞素组装成兔红细胞和人多形核白细胞上的成孔环形复合物” Biosci.Biotechnol.Biochem.63・已出版 5 期（1999 年）。

Kaneko, J.et al.: "An N-terminal region of LukF of staphylococcal leuko-cidin/gamma-hamolysin crucial for the biological activity of the toxin" Biosci.Biotechnol.Biochem.62-7. 1465-1467 (1998)

Kaneko, J. 等人：“葡萄球菌白细胞杀素/γ-单溶素 LukF 的 N 末端区域对于毒素的生物活性至关重要”Biosci.Biotechnol.Biochem.62-7。

Kaneko, J., Mascamas, M.A., Huda, N., Tomita, T., and Kamio, Y.: "An N-terminal region of LukF of staphylococcal leukocidin/γ-hamolysin crucial for the biological activity of the toxin" Biosci.Biotech.Biochem.62・7. 1465-1467 (1998)

Kaneko, J.、Mascamas, M.A.、Huda, N.、Tomita, T. 和 Kamio, Y.：“葡萄球菌杀白细胞素/γ-溶血素的 LukF 的 N 末端区域对于毒素的生物活性至关重要”Biosci .生物技术.生物化学.62・7.

Sugawara, N.et al.: "Assembly of staphylococcal leukocidin into a pore-forming ring-shaped complex on rabbit erythrocytes and human polymorphonuclear leukocytes" Biosci.Biotechnol.Biochem.63-5, (in press, ). (1999)

Sukawara, N.等人：“将葡萄球菌杀白细胞素组装成兔红细胞和人多形核白细胞上的成孔环形复合物”Biosci.Biotechnol.Biochem.63-5，（出版中）。