Roles of ovalbumin on formation of central nerve system in chick embryo

卵清蛋白对鸡胚中枢神经系统形成的作用

• 批准号: 16580240
• 负责人: SUGIMOTO Yasushi
• 金额: $ 2.43万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16580240/
• 关键词: ovalbumin; S-ovalbumin; serpin; gene expression; chick embryo; morphogenic factors; protein structure; NTD; 形態形成; 中枢神経; 神経管欠損; 神経管形成; NTD

Ovalbumin has been classified as a homolog of serine protease inhibitor (serpin), but less is known for its function except for a possible contribution as a nutrient source. We have shown that, during embryonic development, native molecules of ovalbumin (N-ovalbumin) are incorporated into tissues without degradation, while undergoing configurational changes into a heat-stable form named 'HS-cvalbumin. ' Here, to investigate the indispensability of HS-ovalbumin, we established in vitro culture conditions, under which chick embryos, isolated from eggs at stage 7, when closure of neural tube begins, could develop normally for up to 24 hours. When endogenous, remnant ovalbumin was blocked immunologically, the isolated embryos became irregular particularly in the brain tissue, hardly forming the neural tube closure which would occur in normal embryos at stage 9. The failure of closure of neural tube is so-called neural tube defects (NTD). The neural tube of the embryo treated with ovalbumin-antibody had less extremely apoptotic cells and proliferating cells. The abnormal embryos were rescued by re-adding a sufficient amount of HS-ovalbumin to the medium after treating anti-ovalbumin IgG. HS-ovalbumin showed a higher rescue rate compared not only to N- ovalbumin but also to another heat stable counterpart (S-ovalbumin) known to occur artificially by heating N-ovalbumin in vitro. In NTD embryo, expression of FGF8 and Slug has decreased extremely, while Pax 3 up-regulated 5 folds. Many morphogenic and cell cyclic factors were not affected by depletion of ovalbumin. These findings may be the first indication that ovalbumin is an essential protein involved in the normal organogenesis.

卵清蛋白被归类为丝氨酸蛋白酶抑制剂（sereprotein inhibitor，serpin）的同系物，但除了作为营养源的可能贡献外，对其功能知之甚少。我们已经表明，在胚胎发育期间，卵清蛋白（N-卵清蛋白）的天然分子被掺入组织中而不降解，同时经历构型变化成为称为“HS-卵白蛋白”的热稳定形式。“在这里，为了研究HS-卵清蛋白的不可或缺性，我们建立了体外培养条件，在这种条件下，从第7阶段的鸡蛋中分离出来的鸡胚，当神经管开始关闭时，可以正常发育长达24小时。当内源性残留卵清蛋白被免疫学阻断时，分离的胚胎变得不规则，特别是在脑组织中，几乎不形成在第9阶段正常胚胎中会发生的神经管闭合。神经管闭合失败即所谓的神经管缺陷（NTD）。卵清蛋白抗体处理的胚胎神经管有较少的极端凋亡细胞和增殖细胞。在处理抗卵清蛋白IgG后，通过向培养基中再加入足够量的HS-卵清蛋白来挽救异常胚胎。HS-卵白蛋白不仅与N-卵白蛋白相比，而且与已知通过在体外加热N-卵白蛋白而人工发生的另一种热稳定对应物（S-卵白蛋白）相比，显示出更高的拯救率。在NTD胚胎中，FGF 8和Slug的表达显著降低，而Pax 3的表达上调了5倍。许多形态发生和细胞周期因子不受卵清蛋白耗竭的影响。这些发现可能是第一个迹象表明，卵清蛋白是一个重要的蛋白质参与正常器官的发生。

项目成果

Perchioric acid-soluble protein regulates cell proliferation and differentiation in the spinal cord of chick embryos

高氯酸可溶性蛋白调节鸡胚脊髓细胞增殖和分化

• 发表时间: 2005
• 期刊: FEBS Lett. 579(11)
• 作者: Himeno E;Sugimoto Y;Oka T. et al.
• 通讯作者: Oka T. et al.

Improvement of functional properties of egg white protein through phosphorylation by dry-heating in the presence of pyrophosphate

• DOI: 10.1021/jf0498259
• 发表时间: 2004-09-08
• 期刊: JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
• 影响因子: 6.1
• 作者: Li, CP;Ibrahim, HR;Aoki, T
• 通讯作者: Aoki, T

Phosphorylation of ovalbumin by dry-heating in the presence of pyrophosphate: effect on protein structure and some properties.

• DOI: 10.1021/jf047793j
• 发表时间: 2005-05
• 期刊: Journal of agricultural and food chemistry
• 影响因子: 6.1
• 作者: Can-Peng Li;Y. Hayashi;H. Shinohara;H. Ibrahim;Y. Sugimoto;Junichi Kurawaki;N. Matsudomi;T. Aoki-T.-Ao

ベーシックマスター分子生物学

分子生物学基础硕士

• 发表时间: 2007
• 作者: 東中川徹;他
• 通讯作者: 他

Transition of ovalbumin to thermostable structure entails conformational changes the reactive center loop.

卵清蛋白向热稳定结构的转变需要反应中心环的构象变化。

• 发表时间: 2007
• 期刊: Biochim Biophys Acta 1770(1)
• 作者: Shinohara H;Horiuchi M;Sato M;Kurisaki J.Kusakabe T;Koga K;Minami Y;Aoki T;Kato I;Sugimoto Y.
• 通讯作者: Sugimoto Y.