Physiological early response in tissue repair and regeneration -molecular mechanism of shedding : Analyses in wound healing model

组织修复和再生的生理早期反应-脱落的分子机制：伤口愈合模型分析

• 批准号: 13670139
• 负责人: HIGASHIYAMA Shigeki
• 金额: $ 2.62万
• 依托单位: Ehime University (2002)Osaka University (2001)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-13670139/
• 关键词: HB-EGF; shedding; ADAM; wound healing; EGF family; SH3 protein; keratinocyte; metalloprotease; Shedding; Angiotensin II; IL-8; SH3蛋白質; EGFファミリー; PACSIN3; Wound Healing; Metalloprotease

1) TPA-dependent HB-EGF shedding was completely abrogated in ADAM12-/- mouse-derived embryonic fibroblast, but not in ADAM9-/- mouse-derived embryonic fibroblast, suggesting that ADAM12 is a key enzyme in HB-EGF shedding.2) In order to analyze the activation mechanism of ADAM12, we screened proteins to bind the cytoplasmic of ADAM12 using a yeast two-hybrid method, resulting in the identification of two kinds of SH3 domain-containing proteins, PACSIN3 and a novel one designated Eve-1. PACSIN3 consists of 416 amino acids and has three SH3 domains in the C-terminal region. Eve-1 has two spliced isoforms, Eve-1a and Eve-1b which consists 767 and 790 amino acids, respectively. Eve-1a and Eve-1b have four and five SH3 domains in the C-terminal region. Overexpression of PACSIN3 and Eve-1a suppressed additively TPA-HB-EGF shedding.Based on these data, we speculated that HB-EGF shedding would be regulated multiply by ADAM members and SH3 domain containing proteins.

1) TPA 依赖性 HB-EGF 脱落在 ADAM12-/- 小鼠胚胎成纤维细胞中完全消除，但在 ADAM9-/- 小鼠胚胎成纤维细胞中则没有，表明 ADAM12 是 HB-EGF 脱落的关键酶。 2 ）为了分析ADAM12的激活机制，我们利用酵母二杂交法筛选了与ADAM12胞浆结合的蛋白，得到了鉴定了两种含有 SH3 结构域的蛋白质：PACSIN3 和一种名为 Eve-1 的新型蛋白质。 PACSIN3 由 416 个氨基酸组成，C 端区域有 3 个 SH3 结构域。 Eve-1有两种剪接异构体，Eve-1a和Eve-1b，分别由767个和790个氨基酸组成。 Eve-1a 和 Eve-1b 在 C 端区域有 4 个和 5 个 SH3 结构域。 PACSIN3和Eve-1a的过表达抑制了TPA-HB-EGF脱落。基于这些数据，我们推测HB-EGF脱落将受到ADAM成员和含有SH3结构域的蛋白质的多重调节。

项目成果

Kurisaki, T. et al.: "Phenotypic Analysis of Meltrin a (ADAM12) -Deficient Mice: Involvement of Meltrin a in Adipogenesis and Myogenesis"Mol. Cell. Biol.. 23. 55-61 (2003)

Kurisaki, T. 等人：“Meltrin a (ADAM12) 缺陷小鼠的表型分析：Meltrin a 参与脂肪生成和肌生成”Mol。

Asakura, M. et al.: "Cardiac hypertrophy is inhibited by antagonism of ADAM12 processing of HB-EGF: Metalloproteinase inhibitors as a new therapy"Nature Med.. 8. 35-40 (2002)

Asakura, M. 等人：“HB-EGF 的 ADAM12 加工的拮抗作用可抑制心脏肥大：作为新疗法的金属蛋白酶抑制剂”Nature Med.. 8. 35-40 (2002)

Umeda, Y.: "Involvement of heparin-binding EGF-like growth factor and its processing by metalloproteases in early epithelial morphogenesis of the submandibular"Develop. Biol.. 237. 202-211 (2001)

Umeda, Y.：“肝素结合 EGF 样生长因子的参与及其在下颌下早期上皮形态发生中的金属蛋白酶加工”发展。

Yoshinaka, T., Nishii, K., Nishiwaki, E., Yamada, K., Sawada, H., Yoshino, K., Ishiguro, H. and Higashiyama, S.: "Identification and characterization of novel mouse and human ADAM33s with potential metalloprotease activity"Gene. 282. 227-236 (2002)

Yoshinaka, T.、Nishii, K.、Nishiwaki, E.、Yamada, K.、Sawada, H.、Yoshino, K.、Ishiguro, H. 和 Higashiyama, S.：“新型小鼠和人类 ADAM33 的鉴定和表征

Deepa S.S., Higashlyama, S., et al.: "Specific molecular interactions of oversulfated chondroitin sulfate E with various heparin-binding growth factors: Implications as a physiological binding partner in the brain and other tissues."J. Biol Chem.. 277. 43

Deepa S.S.、Higashlyama, S. 等人：“过硫酸化硫酸软骨素 E 与各种肝素结合生长因子的特定分子相互作用：作为大脑和其他组织中的生理结合伴侣的影响。”J.