Development of Sensor System to Analyse and Diagnose the abnormality of Protein Conformation

蛋白质构象异常分析诊断传感器系统的开发

基本信息

批准号：
15206089
负责人：
KUBOI Ryoichi
金额：
$ 27.96万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

The goal of this research project is to establish the analytical method of dynamic interaction between the structurally abnormal protein and biomembrane under the stress condition, especially focusing on the local hydrophobicity (LH) or structural fluctuation. Amyloid beta peptide (Aβ) has been employed as a case study of the system to analyze and characterize the structural abnormality of various proteins. During past two years, we have developed the sensor system of the structural abnormality of protein and have established the database on the protein abnormality under the stress condition. In 2005, the method to judge the structural abnormality of proteins has been investigated based on the previous findings. Based on the database, the suitable conditions (stress condition and lipid compositions) to regenerate the conformational abnormality of protein under the stress condition on the liposome surface have been clarified, resulting that the microdomain-like structure of the membrane … More surface was found to play an important role on the conformational abnormality of proteins/peptides. It was furthermore found that the microdomain-like structure could suitably be controlled by the control of the stress conditions (heating, pH and ionic strength) and the modification of membrane components (alcohols, fatty acids, cholesterol). Especially in the case of the cholesterol-modified liposome, the Aβ-membrane interaction was found to be controlled on the membrane in the presence of metal ion, Cu. The Aβ-Cu complex furthermore induced the metalloenzyme-like function(oxidation of cholesterol and dopamine). The above potential functions of Aβ on the membrane was found to be closely related to the stability of hydrogen-bond of the main chain of the Aβ in the hydrophobic nano environment inside the liposome membrane. By using the membrane chip arraying various stressed-liposomes on the chip, it was found that we could assess the structural abnormality of the protein. The above findings on the structural abnormality of protein were summarized together with the physiological role of amyloid fibril formation of Aβ on the biomembrane. Less

该研究项目的目标是建立结构异常蛋白质与生物膜在应激条件下的动态相互作用的分析方法，特别关注局部疏水性（LH）或淀粉样β肽（Aβ）作为分析方法。分析和表征各种蛋白质结构异常的系统的案例研究在过去的两年里，我们开发了蛋白质结构异常的传感器系统，并建立了应激条件下蛋白质异常的数据库。 2005年，在前人研究成果的基础上，研究了判断蛋白质结构异常的方法，以数据库为基础，研究了脂质体表面在应激条件下再生蛋白质构象异常的适宜条件（应激条件和脂质成分）。已被阐明，结果发现膜表面的微结构域对蛋白质/肽的构象异常起着重要作用，并进一步发现微结构域可以在蛋白质/肽的构象异常中发挥重要作用。通过控制应激条件（加热、pH 和离子强度）和膜成分（醇、脂肪酸、胆固醇）的修饰来适当地控制，特别是在胆固醇修饰的脂质体的情况下，Aβ-膜相互作用受到影响。发现在金属离子 Cu 存在的情况下，Aβ-Cu 复合物进一步诱导了金属酶样功能（胆固醇和多巴胺的氧化）。通过使用在芯片上排列各种应力脂质体的膜芯片，我们发现膜与脂质体膜内疏水性纳米环境中Aβ主链氢键的稳定性密切相关。可以评估蛋白质的结构异常。以上关于蛋白质结构异常的发现与Aβ在生物膜上淀粉样原纤维形成的生理作用进行了总结。