Molecular Anatomy on Intercellular Junctions

细胞间连接的分子解剖学

• 批准号: 03404015
• 负责人: TSUKITA Shigenobu
• 金额: $ 16.32万
• 依托单位: National Institute for Physiological Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03404015/
• 关键词: cell adhesion; adherens junctions; radixin; undercoat; adhesion apparatus; tyrosin phosporylation; tyrosin kinase; monoclonal antibody; アドヘレンス ジャンクション; ラディキシン; 裏打ち蛋白質; チロシリン酸化; モノクローナル〓〓; 細胞接着; カドヘリン; アクチン; アンキリン; カラニン; チロシンキナ-ゼ; カテニン

The ERM family members, ezrin, radixin, and moesin, localizing just beneath the plasma membranes are thought to be involved in the actin filament/plasma membrane association. To identify the intergral membrane protein directly associated with ERM family members, we performed immunoprecipitation studies using anti-moesin mAb and cultured BHK cells metabolically-labeled with [^<35>S]metionine or surface-labeled with biotin. The results indicated that moesin is directly associated with a 140kD integral membrane protein. Using BHK cells as antigens, we obtained a mAb which recognized the 140kD membrane protein. We next cloned a cDNA encoding the 140kD membrane protein and identified it as CD44, a broadly-distributed cell surface glycoprotein. Immunoprecipitation with various anti-CD44 mAbs showed that ezrin and radixin as well as moesin are associated with CD44 not only in BHK cells but also in mouse L fibroblasts. Furthermore, immunofluorescence microscopy revealed that in both BHK and L cells, the Triton X-100-insoluble CD44 is precisely colocalized with ERM family members. We concluded that ERM family members work as molecular linkers between the cytoplasmic domain of CD44 and actin-based cytoskeletons.

ERM家族成员ezrin、radixin和膜突蛋白（moesin）位于质膜下方，被认为参与肌动蛋白丝/质膜缔合。为了鉴定与ERM家族成员直接相关的整合膜蛋白，我们使用抗膜突蛋白mAb和培养的BHK细胞进行免疫沉淀研究，所述细胞用[^<35>S]蛋氨酸代谢标记或用生物素表面标记。结果表明，膜突蛋白与一个分子量为140 kD的膜整合蛋白直接结合。以BHK细胞为抗原，获得了识别140 kD膜蛋白的单克隆抗体。我们随后克隆了编码140 kD膜蛋白的cDNA，并将其鉴定为CD 44，一种广泛分布的细胞表面糖蛋白。免疫沉淀与各种抗CD 44单克隆抗体表明，ezrin和radixin以及膜突蛋白与CD 44不仅在BHK细胞，而且在小鼠L成纤维细胞。此外，免疫荧光显微镜显示，在BHK和L细胞中，Triton X-100不溶性CD 44与ERM家族成员精确共定位。我们的结论是，ERM家族成员作为分子之间的CD 44和肌动蛋白为基础的细胞骨架的胞质结构域的接头。

项目成果

Tsukita,Sa.: "ERM family members as molecular linkers between the cell surface glycoprotein CD44 and actin filaments." J.Cell Biol.(in press). (1994)

Tsukita,Sa.：“ERM 家族成员作为细胞表面糖蛋白 CD44 和肌动蛋白丝之间的分子连接物。”

Tukita, S., Tsukita, S., Nagafuchi, A., and Yonemura, S.: "Molecular linkage between cadherins and actin filaments in cell-cell adherens junctions" Curr. Opin. Cell Biol.4. 834-839 (1992)

Tukita, S.、Tsukita, S.、Nagafuchi, A. 和 Yonemura, S.：“细胞-细胞粘附连接中钙粘蛋白和肌动蛋白丝之间的分子连接”。

Funayama, N., Nagafuchi, A., Sato, N., Tsukita, S., and Tsukita, S.: "Radixin is a novel member of the band 4.1 family." J.Cell Biol.1. 15. 1039-1048 (1991)

Funayama, N.、Nagafuchi, A.、Sato, N.、Tsukita, S. 和 Tsukita, S.：“Radixin 是乐队 4.1 家族的新成员。”

Sato, N., Yonemura, S., Obinata, T., Tsukita, S., and Tsukita, S: "Radixin, a barbed-end-capping actin-modulating protein, is concentrated at the cleavage furrow during cytokinesis" J.Cell Biol.113. 321-330 (1991)

Sato, N.、Yonemura, S.、Obinata, T.、Tsukita, S. 和 Tsukita, S：“Radixin，一种带倒刺末端加帽的肌动蛋白调节蛋白，在胞质分裂过程中集中在卵裂沟” J.

Hashimoto,T.: "Desmoyokin,a 680KDa keratinocyte plasma membraneassociated protein,is homologous to the protein ecoded by AHNAK." J.Cell Sci.105. 275-286 (1993)

Hashimoto,T.：“Desmoyokin，一种 680KDa 的角质形成细胞质膜相关蛋白，与 AHNAK 编码的蛋白同源。”