Analysis of Intermedilysin in Human Specific Cytolytic Mechanism : Design and Application for Cell Targeting Module.

人特异性溶细胞机制中中间素的分析：细胞靶向模块的设计和应用。

• 批准号: 12672151
• 负责人: OHKURA Kazuto
• 金额: $ 2.18万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12672151/
• 关键词: cytolysin; targeting; infection; コレステロール

Intermedilysin (ILY), a human specific cytolysin secreted by Streptococcus intermedius, is a member of the gene of SLO-family cytolysin (SFC), but differs by exhibiting a low affinity to cholesterol (CHL) in addition to human specificity. We modeled ILY molecule using perfringolysin O (PFO) X-ray data, and observed four domain (domain 1 〜 domain 4) in the molecule. From the analysis of ILY model, the domain 4 seems to be the human cell membrane associated region. Especially, extended-NTD sequence at ILY C-terminal appeared an important factor for human specific cytolysis. Indeed, the NTD-deleted ILY mutant (ΔNTD-ILY) decreased the hemolytic activity for human erythrocytes. Then we considered that the C-terminal NTD should be concerning with the cluster formation on the human cell membrane.Moreover we constructed the antiCEA-antibody conjugated ILY domain4 (antiCEA-ILY4D), and examined the cell delivery system. AntiCEA-ILY4D bind to human erythrocyte (ET) membrane easily, and this ET-AntiCEA-ILY4D module recognized CEA-positive TT cell. Now we are improving these targeting modules.

中间体溶素（intermediolysin，ILY）是由中间链球菌分泌的一种人特异性溶细胞素，属于SLO家族溶细胞素（SFC）基因成员，但与人特异性溶细胞素不同，ILY对胆固醇（CHL）的亲和力较低。我们利用产气荚膜梭菌溶素O（PFO）的X射线数据对ILY分子进行了建模，并观察到分子中的四个结构域（结构域1和结构域4）。从ILY模型分析，结构域4可能是人细胞膜相关区。特别是ILY C端的延伸NTD序列是导致人特异性细胞溶解的重要因素。事实上，NTD缺失的ILY突变体（ΔNTD-ILY）降低了对人红细胞的溶血活性。我们进一步构建了抗CEA抗体结合ILY结构域4（antiCEA ILY 4D），并对其细胞传递系统进行了研究。AntiCEA-ILY 4D与人红细胞（ET）膜容易结合，该ET-AntiCEA-ILY 4D模块识别CEA阳性TT细胞。现在我们正在改进这些瞄准模块。

项目成果

Hori, H.: "TX-1123 : An Antitumor 2-Hydroxyaryliden-4-cyclopentene-1, 3-Dione as a Protein Tyrosine Kinase Inhibitor Having Low Mitochondrial Toxicity"Bioorg.Med.Chem.. 10. 3257-3265 (2002)

Hori，H.：“TX-1123：作为具有低线粒体毒性的蛋白酪氨酸激酶抑制剂的抗肿瘤 2-Hydroxyaryliden-4-cyclopentene-1, 3-Dione”Bioorg.Med.Chem.. 10. 3257-3265 (2002

Goto,T., Nagamune,N., Kawamura,Y., Ohnishi,O., Hattori,K., Ohkura,K., Miyamoto.K., Akimoto,S., Ezaki,T., Hirota,K., Miyake,Y., Maeda,T.and Kourai,H.: "Rapid Identification of Streptococcus intermedius by PCR using the ily gene as aspecies marker gene"J.Me

后藤，T.，长宗，N.，川村，Y.，大西，O.，服部，K.，大仓，K.，宫本，K.，秋元，S.，江崎，T.，广田，K.，

Kasai,S.: "New Antimetastatic Hypoxic Cell Radiosensitizer : Design, Synthesis, and Biological Activities of 2-Nitroimidazole-acetamide, TX-1877, and Its Analogues."Bioorg.Med.Chem.. in press. (2001)

Kasai,S.：“新型抗转移缺氧细胞放射增敏剂：2-硝基咪唑-乙酰胺、TX-1877 及其类似物的设计、合成和生物活性。”Bioorg.Med.Chem.. 正在出版。

Ohkura, K.: "Modification of Cell Response to Insulin by Membrane-Acting Agents in Rat White Adipocytes Analysis of Structural Features by Computational Simulation"Bioorg.Med.Chem.. 9. 3023-3033 (2001)

Ohkura, K.：“通过计算模拟分析大鼠白色脂肪细胞中的膜作用剂对胰岛素的细胞反应的结构特征分析”Bioorg.Med.Chem.. 9. 3023-3033 (2001)

Ohkura,K.and Hori,H.: "Modification of Cell Response to Insulin by Membrane-Acting Agents in Rat White Adipocytes : Analysis of Structural Features by Computational Simulation"Bioorg.Med.Chem.. 9. 3023-3033 (2001)

Ohkura,K. 和 Hori,H.：“大鼠白色脂肪细胞中膜作用剂对胰岛素的细胞反应的修改：通过计算模拟分析结构特征”Bioorg.Med.Chem.. 9. 3023-3033 (2001)