CLAUDINS AND OCCLUDIN : COMPARISON WITH CONNEXIN

CLAUDINS 和 OCCLUDIN：与 CONNEXIN 的比较

• 批准号: 11694270
• 负责人: TSUKITA Shoichiro
• 金额: $ 4.16万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694270/
• 关键词: OCCLUDIN; CLAUDIN; CONNEXIN; TIGHT JUNCTION; GAP JUNCTION; CHIMERIC MOLECULE; CRYSTALIZATION; GENE FAMILY; ギャップジャンクション; 膜蛋白質; 結晶化; 電子顕微鏡

In tight junctions (TJs), TJ strands are associated laterally with those of adjacent cells to form paired strands to eliminate the extracellular space. Claudin-1 and-2, integral membrane proteins of TJs, reconstitute paired TJ strands when transfected into L fibroblasts. Claudins comprise a multigene family and more than two distinct claudins are co-exrressed in single cells, raising the questions of whether heteroseneous claudins form heteromeric TJ strands and whether claudins interact between each of the paired strands in a heterophilic manner. To answer these questions, we co-transfected two of claudin-1, -2 and -3 into L cells, and detected their coconcentration at cell-cell borders as elaborate networks. Immunoreplica electron microscopy confirmed that distinct claudins were co-incorporated into individual TJ strands. Next, two L transfectants singly expressing claudin-1, -2 or -3 were cocultured, and we found that claudin-3-strands laterally associated with claudin-1 and claudin-2 stranas to form paired strands, whereas claudin-1 strands did not interact with claudin-2 strands. We concluded that distinct species of claudins can interact within and between TJ strands except in some combinations. This mode of assembly of claudins could increase the diversity of the structure and functions of TJ strands.

在紧密连接（TJ）中，TJ链与相邻细胞的侧面相关联，以形成成对的链，以消除细胞外空间。 Claudin-1和-2，TJ的整体膜蛋白，当转染成L成纤维细胞时，重建成对的TJ链。 Claudins包括一个多基因家族和两个以上不同的Claudins在单个细胞中共脱离，这提出了异质claudins是否形成异源TJ链的问题，以及Claudins是否以异性词的方式在每个配对链之间相互作用。为了回答这些问题，我们将Claudin-1，-2和-3的两个共转染到L细胞中，并在细胞细胞边界中检测到它们作为精美的网络。 Immunoreplica电子显微镜证实，将不同的Claudins共同纳入单个TJ链中。接下来，将两个L转染剂单独表达claudin-1，-2或-3被共培养，我们发现与Claudin-1和Claudin-1和Claudin-2 Stranas横向相关的Claudin-3链形成了成对的链，而Claudin-1链没有与Claudin-2链相互作用。我们得出的结论是，除了某些组合外，不同种类的claudins可以在TJ链之间和之间相互作用。这种claudins组装方式可以增加TJ链的结构和功能的多样性。

项目成果

Morita.K, Sasaki, H., Fujimoto, K., Furuse, M., and Tsukita, Sh.: "Claudin-11/OSP-based tight junctions in myelinated sheaths of oligodendrocytes and in Sertoli cells in testis."J.Cell Biol.. 145. 579-588 (1999)

Morita.K、Sasaki, H.、Fujimoto, K.、Furuse, M. 和 Tsukita, Sh.：“少突胶质细胞有髓鞘和睾丸支持细胞中基于 Claudin-11/OSP 的紧密连接。”

斎藤通紀: "Complex phenotype of mice lacking occludin, a component of tight junction"Molecular Biology of the Cell. 11. 4131-4142 (2000)

Michinori Saito：“缺乏紧密连接成分occludin 的小鼠的复杂表型”《细胞分子生物学》11. 4131-4142 (2000)。

月田承一郎: "Pores tin the wall : Claudins constitute tight junction strands containing aqueous pores"Journal of Cell Biology. 149. 13-16 (2000)

Joichiro Tsukita：“壁上的孔：Claudins 构成含有水孔的紧密连接链”，细胞生物学杂志 149. 13-16 (2000)。

Itoh, M., Furuse, M., Morita, K., Kubota, K., and Tsukita, Sh.: "Molecular basis for the recruitment of there MAGUKs. ZO-1, ZO-2 and ZO-3, to tight junctions : Direct binding of their first PDZ domains to the COOH-termini of claudins."J.Cell Biol.. 147. 1

Itoh, M.、Furuse, M.、Morita, K.、Kubota, K. 和 Tsukita, Sh.：“招募 MAGUK 的分子基础。ZO-1、ZO-2 和 ZO-3，以紧密

古瀬幹夫: "Conversion of Zonula Occludentes from tight to leaky strand type by introducting claudin-2 into MDCK I cells"Journal of Cell Biology. (印刷中). (2001)

Mikio Furuse：“通过将claudin-2 引入 MDCK I 细胞将闭锁带从紧密链型转变为渗漏链型”，《细胞生物学杂志》（2001 年出版）。