Establishment of Ryudocan Null Mouse and ELISA for Blood Levels of Ryudocan

Ryudocan 空鼠的建立及 Ryudocan 血药浓度的 ELISA 测定

• 批准号: 10557090
• 负责人: KOJIMA Tetsuhito
• 金额: $ 8.45万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10557090/
• 关键词: Heparan sulfate proteoglycan; ryudocan (syndecan-4); null mice; focal adhesion; fibronectin; Ryudocan; ノックアウトマウス; 血中濃度; 心筋梗塞; 組織修復

Focal adhesion formation on fibronectin requires two signals, i.e. one from the cell-binding domain of fibronectin and one from the heparin-binding domain. While the cell-binding domain is recognized by integrins, the cell surface molecules that recognize the heparin-binding domain have not been identified. Previous reports have demonstrated that ryudocan (syndecan-4), a heparan sulfate proteoglycan, is a component of focal adhesions and that anti-syndecan-4 antibody can promote focal adhesion formation. Thus, ryudocan is a candidate that recognizes the heparin-binding domain. To study the role of syndecan-4 in focal adhesion formation, we generated ryudocan null mice. Even in ryudocan (-/-) fibroblasts, focal adhesions were formed and actin fibers terminated normally at focal adhesions. However, neither the fibronectin heparin-binding fragment in a soluble form nor anti-ryudocan antibody promoted focal adhesion formation in ryudocan (-/-) fibroblasts cultured on the cell-binding fragment, promoted focal adhesion formation. Thus, the present study has revealed that the heparin-binding fragment in a soluble form requires ryudocan for focal adhesion formation, whereas one in a solid form could utilize not only ryudocan but also some unidentified cell surface molecules.

纤连蛋白上的粘着斑形成需要两种信号，即一种来自纤连蛋白的细胞结合结构域，一种来自肝素结合结构域。虽然细胞结合结构域被整联蛋白识别，但识别肝素结合结构域的细胞表面分子尚未鉴定。之前的报道表明，ryudocan（syndecan-4）是一种硫酸乙酰肝素蛋白聚糖，是粘着斑的组成部分，并且抗syndecan-4抗体可以促进粘着斑形成。因此，ryudocan 是识别肝素结合域的候选者。为了研究 syndecan-4 在粘着斑形成中的作用，我们生成了 ryudocan null 小鼠。即使在ryudocan (-/-)成纤维细胞中，也形成粘着斑并且肌动蛋白纤维在粘着斑处正常终止。然而，可溶形式的纤连蛋白肝素结合片段和抗ryudocan抗体均不促进在细胞结合片段上培养的ryudocan(-/-)成纤维细胞中的粘着斑形成，促进粘着斑形成。因此，本研究表明，可溶形式的肝素结合片段需要ryudocan来形成粘着斑，而固体形式的肝素结合片段不仅可以利用ryudocan，还可以利用一些未鉴定的细胞表面分子。

项目成果

A. Katsumi, T. Kojima, T. Yamazaki, et al.: "The Carboxyl-Terminal Region of Protein C is Essential for Its Secretion"Blood. 91. 3784-3791 (1998)

A. Katsumi、T. Kojima、T. Yamazaki 等人：“C 蛋白的羧基末端区域对于其分泌至关重要”血液。

K.Ishiguro, T.Kojima, O.Taguchi, et al.: "Syndecan-4 expression is associated with follicular atresia in mouse ovary."Histochem. Cell Biol.. 112. 25-33 (1999)

K.Ishiguro、T.Kojima、O.Taguchi 等人：“Syndecan-4 表达与小鼠卵巢中的卵泡闭锁有关。”Histochem。

T.Nakanishi,K.Kadomatsu,T.Kojima,et al.: "Expression of Syndecan-1 and -3 during Embryogenesis of the Central Nerve System in Relation to Binding with Midkine." J.Biochem.121. 197-205 (1997)

T.Nakanishi、K.Kadomatsu、T.Kojima 等人：“中枢神经系统胚胎发生过程中 Syndecan-1 和 -3 的表达与中期因子结合的关系”。

K. Ishiguro, K. Kadomatsu, T. Kojima, et al.: "Syndecan-4 Deficiency Impairs Focal Adhesion Formation Only under Restricted Conditions"J. Biol. Chem.. 275. 5249-5252 (2000)

K. Ishiguro、K. Kadomatsu、T. Kojima 等人：“Syndecan-4 缺陷仅在限制条件下会损害局部粘连形成”J.

A. Shimizu, I. Sugiura, T. Kojima, et al.: "Identification of the Five Hydrophilic Residues (Lys-217, Lys-218, Arg-359, His-360, and Arg-513) Essential for the Structure and Activity of Vitamin K-Dependent Carboxylase"Biochem. Biophys. Res. Commn.. 251. 2

A. Shimizu、I. Sugiura、T. Kojima 等人：“鉴定对结构和结构至关重要的五种亲水残基（Lys-217、Lys-218、Arg-359、His-360 和 Arg-513）