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.

外结构域脱落是肝素结合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控。然而，胞外结构域脱落的分子机制在很大程度上仍有待阐明。结果表明：1)在Vero细胞中，金属蛋白酶结构域缺陷型ADAM12突变体的过表达诱导HB-EGF外结构域脱落。几种ADAM12突变体的过表达和抗ADAM12标签抗体的共沉淀实验表明，ADAM12可能作为信号转导的适配蛋白，而不是作为蛋白酶，促进HB-EGF脱落。2)筛选具有抑制HB-EGF外畴脱落活性的低分子量化学物质。少数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，构建了包含8.4 k人类基因组序列的siRNA文库，并利用逆转录病毒载体系统转染人细胞。对脱落刺激无反应的细胞群被分离。

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.