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TJ链横向相连，形成成对的链，以消除细胞外间隙。紧密连接蛋白-1和-2，完整的膜蛋白的TJ，重组配对的TJ链转染到L成纤维细胞时。Claudins包括一个多基因家族和两个以上的不同的claudins在单细胞中共表达，提出的问题是否heteroseneous claudins形成异聚TJ链和是否claudins之间的相互作用的每对链之间的heterophilic方式。为了回答这些问题，我们将claudin-1、claudin-2和claudin-3中的两种共转染到L细胞中，并检测它们在细胞-细胞边界处的共浓度作为精细的网络。Immunoreplica电子显微镜证实，不同的claudins共同纳入个别TJ链。接下来，将单独表达claudin-1、claudin-2或claudin-3的两个L转染子共培养，并且我们发现claudin-3-链与claudin-1和claudin-2链横向缔合以形成成对链，而claudin-1链不与claudin-2链相互作用。我们的结论是，不同种类的claudins可以相互作用的TJ链内和之间，除了在某些组合。这种紧密连接蛋白的组装模式可以增加TJ链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)。

Furuse, M., Sasaki.H., and Tsukita, Sh.: "Manner of interaction of heterogenous claudin species within and between tight junction strands."J.Cell Biol.. 147. 891-903 (1999)

Furuse, M.、Sasaki.H. 和 Tsukita, Sh.：“紧密连接链内和之间的异源密蛋白种类的相互作用方式。”J.Cell Biol.. 147. 891-903 (1999)

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）。

月田承一郎: "Multifunctional strands in tight juncntions"Nature Review Molecular Cell Biology. (印刷中).

Joichiro Tsukita：“紧密连接中的多功能链”《自然评论分子细胞生物学》（正在出版）。