Crystal Structural Analysis of IL6/IL6 receptor Complex and Development of Functional Regulation of IL6 with cDNA

IL6/IL6 受体复合物的晶体结构分析及 cDNA 对 IL6 功能调节的开发

• 批准号: 13359003
• 负责人: TSUKIHARA Tomitake
• 金额: $ 31.78万
• 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: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13359003/
• 关键词: IL6; IL6 recetor; crvstal structure; AFM; signal transduction; インターロイキン-6; インターロイキン-6受容体; 複合体; 結晶構造解析

1. Atotal of 150 mg AUK18l 6 was purified from 30 ml of abdominal dropsy. Sil6 receptor affinity column was prepared by using AUK181 6. 6 mg of Sil6 receptor was purified from CHO cells producing Sil6 receptor.2. Structure of IL 6 in the novel crystal form was determined at 2.3 A resolution. The crystal shucture showed a loop structure that is not visible in the crystal structure previously determined. This loop interacts probably with IL6 receptor.3. Inhibition of IL6 by a FELRY peptide that bind to IL6 was inspected. The FELRY at 0.2 to 25 % saturation was added to IL6 dependent cells under rIL6 stimulation. Significant suppression of cell growth was not detected. Other soluble peptides are in under inspection.4. The method for molecular observation by AFM was developed to analyze recognition among IL6, IL6 receptor and gp130.

1.从30 ml腹水中纯化出150 mg AUK1816。用AUK 181 6制备了Si1 6受体亲和柱。从产生Sil6受体的CHO细胞中纯化6 mg Sil6受体。在2.3A分辨率下测定了新晶体形式的IL 6的结构。晶体结构显示出在先前测定的晶体结构中不可见的环状结构。该环可能与IL 6受体相互作用.检查结合IL 6的FELRY肽对IL 6的抑制。在rIL 6刺激下，将0.2%至25%饱和度的FELRY加入IL 6依赖性细胞中。未检测到细胞生长的显著抑制。其他可溶性肽也在考察中.建立了利用原子力显微镜进行分子观察的方法，分析了IL 6、IL 6受体和gp130之间的识别。

项目成果

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