Structural basis for molecular mechanism of heparanase mediated cancer cell invasion and metastasis

乙酰肝素酶介导癌细胞侵袭和转移分子机制的结构基础

• 批准号: 23790048
• 负责人: TABATA Kaori
• 金额: $ 2.75万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23790048/
• 关键词: ヘパラナーゼ; X線結晶構造解析; ガン転移; 結晶構造解析

Heparanase is an enzyme which involved in cancer cell invasion and metastasis. In this research, the purpose is to clarify the 3D-structure of heparanase.I examined various protein expression systems using E. coli and silk worm, and succeeded in obtaining about 5 mg of the target protein from the 1L culture as a soluble fraction. The antibody was produced using the prepared recombinant protein as an antigen, and I confirmed that the antibody specifically recognized heparanase by ELISA. Then, I tried to crystallize complexes of heparanase with produced antibody, and succeeded in obtaining some micro crystals. Moreover, I also succeeded in obtaining the single crystals of beta-glucuronidase, which belongs to heparanase type enzyme.

肝素酶是一种参与癌细胞侵袭和转移的酶。在这项研究中，目的是阐明肝素酶的3D结构。我使用大肠杆菌和丝绸蠕虫研究了各种蛋白质表达系统，并成功地从1L培养物中获得了约5 mg的靶蛋白作为可溶性馏分。使用制备的重组蛋白作为抗原产生抗体，我证实抗体由ELISA特异性识别的乙par菜酶。然后，我试图通过产生的抗体结晶肝素酶的复合物，并成功获得了一些微晶。此外，我还成功地获得了属于肝素酶类型酶的β-葡萄糖醛酸酶的单晶。

项目成果

β-glucuronidaseの反応メカニズムに関する研究

β-葡萄糖醛酸酶反应机理研究

• 发表时间: 2012
• 作者: 今村聡;今村麻美;坂元政一;田畑香織;田中宏幸;森元聡;コガネバナ
• 通讯作者: コガネバナ