Study on Design of Amphiphilic Functional Peptides and Their Action to Lipid Membranes

两亲性功能肽的设计及其脂膜作用研究

• 批准号: 10680570
• 负责人: AOYAGI Haruhiko
• 金额: $ 2.37万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680570/
• 关键词: amphiphilic peptide; model peptide; phosphilipid membrane; antibacterial activity; lectin; hemolytic activity; nucleic acid carrier; galactose modification; 膜破壊能; コンジュゲート; DNA細胞内移入; アミノ酸置換アナログ; β構造; αヘリシティ

Peptide contributes not only elucidation of action mechanism and structure-function relationship but also to drug design and modeling of functional materials. We have studied the structure-function relationship of antibacterial peptides, pleurocidin (Ple) and bactenecin 5 (Bac 5), effect of chemical modification of hemolytic lectin CEL-III on its cytotoxic activity, and use of cationic amphiphilic peptides as gene carriers into cells. 1) Using some amino acid-substituted Ple analogs, we examined their interaction with lipid and cell membranes. Balance of hydrophobicity and positive charge of Ple was found to be important for antibacterial activity. About Bac 5, it became clear that a certain chain length in the central repeating region is essential for antibacterial activity and the strongly cationic N-terminal portion enhances the activity. 2) By modification of the side chains of Glu and Asp residues in CEL-III with Gly-OMe, not only hemolytic activity but also carbohydrate-binding and agglutinative activities were greatly decreased, suggesting that carboxylic residues at the binding or near site are modified and the binding with carbohydrate would be interrupted. 3) Cationic amphiphilic peptides were modified with galactose, and their DNA transfer ability was evaluated using a human hepatoma cell line. Higher ability was observed with increasing galactose residues. Furthermore, we found that internalization of the DNA-galactose-modified peptide complexes is caused by the receptor-mediated endocytosis.

肽不仅有助于阐明作用机制和结构功能关系，而且有助于药物设计和功能材料的建模。本文研究了抗菌肽pleurocidin（Ple）和bactenecin 5（Bac 5）的结构与功能关系，溶血凝集素CEL-III的化学修饰对其细胞毒活性的影响，以及阳离子两亲肽作为基因载体进入细胞的应用。1)使用一些氨基酸取代的Ple类似物，我们研究了它们与脂质和细胞膜的相互作用。Ple的疏水性和正电荷的平衡被认为是重要的抗菌活性。关于Bac 5，很明显，在中心重复区域中的一定链长对于抗菌活性是必不可少的，并且强阳离子N-末端部分增强活性。2)用Gly-OMe修饰CEL-III中Glu和Asp残基的侧链，不仅溶血活性大大降低，而且糖结合和凝集活性也大大降低，这表明结合位点或附近位点的羧基残基被修饰，与糖的结合将被中断。3)用半乳糖修饰阳离子两亲性肽，并使用人肝癌细胞系评估其DNA转移能力。随着半乳糖残基的增加，观察到更高的能力。此外，我们还发现DNA-半乳糖修饰肽复合物的内化是由受体介导的内吞作用引起的。

项目成果

Takuro Niidome: "Membrane-Perturbing and -Fusogenic Activities of Amphiphilic Helical Peptides and Their Usage in Gene Systems into Cells"Current Topics in Peptide & Protein Res.. 3. 79-89 (1999)

Takuro Niidome：“两亲性螺旋肽的膜扰动和融合活性及其在细胞基因系统中的用途”肽当前主题

Takuro Niidome: "Antibacterial Activity of Arg/Pro-Rich Bactenecin 5 Model Peptides and Their Interaction with Phospholipid Membranes"Bull.Chem.Soc.Jpn.. 73・6. 1397-1402 (2000)

Takuro Niidome：“Arg/Pro-Rich Bactenecin 5 模型肽的抗菌活性及其与磷脂膜的相互作用”Bull.Chem.Soc.Jpn.. 73・6 (2000)。

Takuro Niidome: "Gene Transfer into Hepatoma Cells Mediated by Galactose-Modified α-Helical Peptides"Biomaterials. 21・17. 1811-1819 (2000)

Takuro Niidome：“半乳糖修饰的α-螺旋肽介导的基因转移”生物材料21・17（2000）。

Kazutoshi Yoshida: "Interaction of Pleurocidin and Its Analogs with Phospholipid Membrane and Their Antibacterial Activity"J.Peptide Res.. 57・2. 119-126 (2001)

吉田一俊：“杀侧耳素及其类似物与磷脂膜的相互作用及其抗菌活性”J.Peptide Res. 57・2（2001）。

Kazutoshi Yoshida: "Interaction of Pleurocidin and Its Analogs with Phospholipid Membrane and Their Antibacterial Activity"J.Peptide Res.. (in press). (2001)

Kazutoshi Yoshida：“杀菌素及其类似物与磷脂膜的相互作用及其抗菌活性”J.Peptide Res..（印刷中）。