Analysis of the ectodomain shedding mechanism of HB-EGF

HB-EGF胞外域脱落机制分析

• 批准号: 16207014
• 负责人: MEKADA Eisuke
• 金额: $ 32.7万
• 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: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16207014/
• 关键词: HB-EGF; ectodomain shedding; protease; ADAM; エクトドメインシェディング; シグナル伝達

Ectodomain shedding is critical process for the action of heparin-binding EGF-like growth factor (HB-EGF). This process is regulated by a number of extracellular stimuli and intracellular signaling molecules. However, the molecular mechanism for ectodomain shedding has largely remained to be clarified. We found the following results :1)Over-expression of ADAM12 mutant form with defective metalloprotease domain induced ectodomain shedding of HB-EGF in Vero cells. Over-expression of several mutant forms of ADAM12 and co-precipitation experiments with anti-ADAM12 tag antibody suggested that ADAM12 may contribute to HB-EGF shedding as an adapter protein for signaling, rather than as a protease.2)Screening of low molecular-weight chemicals which possess the inhibitory activity for HB-EGF ectodomain shedding was performed. A few compounds showed the inhibitory activity. Further characterization is underway.3)To understand the molecules which are involved in ectodomain shedding of HB-EGF, we constructed a screening strategy which enables to identify genes necessary for the shedding. To do this, siRNA library containing 8.4 k human genome sequence was constructed and transfected in human cells by retro virus vecter system. Cell populations which did not respond to the stimuli for shedding were isolated.

胞外域脱落是肝素结合 EGF 样生长因子 (HB-EGF) 发挥作用的关键过程。这一过程受到许多细胞外刺激和细胞内信号分子的调节。然而，胞外域脱落的分子机制在很大程度上仍有待阐明。我们发现以下结果：1)具有缺陷金属蛋白酶结构域的ADAM12突变体的过度表达诱导Vero细胞中HB-EGF的胞外域脱落。 ADAM12的几种突变形式的过表达和抗ADAM12标签抗体的共沉淀实验表明ADAM12可能作为信号传导的衔接蛋白而不是作为蛋白酶促进HB-EGF脱落。2)筛选具有HB-EGF胞外域脱落抑制活性的低分子量化学物质。一些化合物显示出抑制活性。进一步的表征正在进行中。3)为了了解参与HB-EGF胞外域脱落的分子，我们构建了一种筛选策略，能够识别脱落所需的基因。为此，构建了含有 8.4 k 人类基因组序列的 siRNA 文库，并通过逆转录病毒载体系统转染人类细胞。分离对脱落刺激没有反应的细胞群。

项目成果

HB-EGF promotes epithelial cell migration in eyelid development

• DOI: 10.1242/dev.02030
• 发表时间: 2005-10-01
• 期刊: DEVELOPMENT
• 影响因子: 4.6
• 作者: Mine, N;Iwamoto, R;Mekada, E
• 通讯作者: Mekada, E

Heparin-binding EGF-like growth factor and the LPA-induced ectodomain shedding pathway is a promising target for the therapy of ovarian cancer.

肝素结合 EGF 样生长因子和 LPA 诱导的胞外域脱落途径是卵巢癌治疗的一个有前景的靶点。

• 发表时间: 2004
• 期刊: Cancer Res. 64・16
• 作者: Miyamoto;S.他12名
• 通讯作者: S.他12名

Suppression of the biological activities of the EGF-like domain by the heparin-binding domain of heparin-binding EGF-like growth factor.

肝素结合 EGF 样生长因子的肝素结合结构域对 EGF 样结构域的生物活性的抑制。

• 发表时间: 2004
• 期刊: J.Biol.Chem. 279・45
• 作者: Takazaki;R.他3名
• 通讯作者: R.他3名

Soluble form of heparin-binding EGF-like growth factor contributes to retinoic acid-induced epidermal hyperplasia

• DOI: 10.1247/csf.30.35
• 发表时间: 2005-12-01
• 期刊: CELL STRUCTURE AND FUNCTION
• 影响因子: 1.5
• 作者: Kimura, R;Iwamoto, R;Mekada, E
• 通讯作者: Mekada, E

Clinical significance of heparin-binding EGF (epidermal growth factor)-like growth factor and ADAM (a disintegrin and metalloprotease) 17 expression in human ovarian cancer.

人卵巢癌中肝素结合 EGF（表皮生长因子）样生长因子和 ADAM（解整合素和金属蛋白酶）17 表达的临床意义。

• 发表时间: 2005
• 期刊: Clin. Cancer Res. 11
• 作者: Tanaka;T.
• 通讯作者: T.