Study on the Membrane-damaging Action of Staphylococcal Alpha-toxin by use of Multilamellar Liposomes.

利用多层脂质体研究葡萄球菌α-毒素的膜损伤作用。

• 批准号: 01570230
• 负责人: TOMITA Toshio
• 金额: $ 1.34万
• 依托单位: The Institute of Medical Science, The University of Tokyo.
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570230/
• 关键词: Staphylococcus aureus; alpha-toxin; membrane damage; liposome; transmembrane channel; membrane fluidity; monoclonal antibody; 毒素六量体形成

Staphylococcal alpha-toxin is a cytolytic 33K-Da polypeptide secreted by pathogenic Staphylococcus aureus. It binds to the membrane of target cells and assembles to hexamer, exhibiting transmembrane-channel activitiy. The toxin is considered to be a prototype for transmembrane-pore-forming proteins including streptolysin O, complement and perforin.By use of multilamellar liposome composed of Phosphatidyl-Choline (PC) as a model membrane, we studied influence of membrane fluidity on the assembly process of the toxin. Our results indicated that alpha-toxin assembled into hexameric form via a transient, nonhemolytic-monomer form, when PC membrane was fluidized either by the phase transition from gel to liquid crystalline state or by the inclusion of >30 mol % cholesterol in the PC membrane.To determine the domain of alpha-toxin which is involved in each step of the assembly process, we prepared monoclonal antibodies to the toxin. Monoclonal antibodies obtained from 14 different hybridoma strains were categorized to 3 groups according to their inhibitory action on binding and hemolytic activity of alpha-toxin. Analysis of the epitope which is recognized by each of the monoclonal antibodies is now in progress.

葡萄球菌α-毒素是一种由金黄色葡萄球菌分泌的细胞溶解33K-DA多肽。它与靶细胞的膜结合并组装与己酰胺，表现出跨膜通道激活。毒素被认为是跨膜孔形成蛋白的原型毒素的过程。我们的结果表明，当PC膜通过从凝胶到液体晶体状态的相变流动或通过将> 30 mol％胆固醇纳入PC膜中时，α-毒素是通过瞬时非溶应能形式组装成六聚体形式的为了确定与组装过程的每个步骤有关的α-毒素的结构域，我们制备了与毒素的单克隆抗体。根据14种不同杂交瘤菌株获得的单克隆抗体，根据其抑制性作用对α-毒素的结合和溶血活性的抑制作用。现在正在进行对每种单克隆抗体识别的表位的分析。

项目成果

Tomita,T.,Watanabe,M.,Takahashi,T.,Kumai,K.,Tadakuma,T.,and Yasuda.T.: "Temperature-sensitive release of adriamycin,an amphiphilic antitumor agent,from dipalmitohlphophatidylcholine-cholesterol liposomes" Biochim.Biophys.Acta. 978. 185-190 (1989)

Tomita,T.、Watanabe,M.、Takahashi,T.、Kumai,K.、Tadakuma,T. 和 Yasuda.T.：“从二棕榈酰磷脂酰胆碱-胆固醇脂质体中温敏释放阿霉素（一种两亲性抗肿瘤剂）”

Yanagie, H., Tomita, T., Sekiguchi, M., and Nariuchi, H.: "Application of boron-10 entrapped in liposomes conjugated with anti-human CEA monoclonal antibody to boron-neutron capture therapy." Brit. J. Cancer Res.

Yanagie, H.、Tomita, T.、Sekiguchi, M. 和 Nariuchi, H.：“将硼 10 包埋在与抗人 CEA 单克隆抗体缀合的脂质体中在硼中子捕获疗法中的应用。”

富田敏夫: "ブドウ球菌α毒素の膜障害機構について" 日本細菌学雑誌. 44. 67 (1989)

Toshio Tomita：“论葡萄球菌α毒素的膜损伤机制”日本细菌学杂志 44. 67 (1989)。

Nakata,M.,Tomita,T.,and Kanegasaki,S.: "Inhibitory effect of antibiotic cerulenin on the respiratory burst in/phagocytes.I.Effects of cerulenin on active oxygen-generation and lipid metabolism" J.Antibiotics. 42. 1171-1177 (1989)

Nakata,M.、Tomita,T. 和 Kanegasaki,S.：“抗生素浅蓝素对吞噬细胞呼吸爆发的抑制作用。I.浅蓝素对活性氧生成和脂质代谢的影响”J.Antibiotics。

Yanagie,H.,Tomita,T.,Sekiguchi,M.,Nariuchi,H.,and Kobayashi,H.: "Application of boronー10 entrapped in liposomes conjugated to antiーhuman CEA monoclonal antibody to boronーneutron capture therapy." Brit.J.Cancer Res.

Yanagie, H.、Tomita, T.、Sekiguchi, M.、Nariuchi, H. 和 Kobayashi, H.：“将硼 10 包埋在与抗人 CEA 单克隆抗体缀合的脂质体中在硼中子捕获疗法中的应用。 “英国癌症研究中心。