Formation of microdomains of the plasma membrane that are involved in intracellular signal transduction

参与细胞内信号转导的质膜微结构域的形成

• 批准号: 13670130
• 负责人: SHIRATAKI Hiromichi
• 金额: $ 2.3万
• 依托单位: DOKKYO UNIVERSITY SCHOOL OF M;EDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670130/
• 关键词: Raft; SNARE; Syntaxin; Taxilin; α-fodrin; Cytoskelton; Coiled-coil domain; SNARE系; 細胞内小胞輸送

Accumulating evidence suggests that formation of microdomains on the plasma membrane is an important process for intracellular signal transduction. Especially, the formation of raft is essential for intracellular signal transduction through GPI-anchored proteins. It has been previously revealed that syntaxin-3, a component of the SNARWE machinery, is implicated in the formation of raft through the regulation of intracellular vesicle transport. However, the molecular mechanism by which the formation of raft is regulated by syntaxin-3 is obscure. Then, in this study, we attempted to identify new syntaxin-3-interacting molecules. We revealed that α-fodrin, a major component of the submembranes cytoskelton, interacted with syntaxin-3. α-Fodrin bound to syntaxin-1 and syntaxin-4 as well as syntaxin-3 but no to syntaxin-7 or -8. α-Fodrin and nc18 or SNAP-25 mutually interacted with syntaxin-1. These results indicate that the formation of the SNARE complex is regulated by α-fodrin and suggest that actin/fodrin meshworks beneath the plasma membrane are involved in the formation of raft. Moreover, we identified a novel syntaxin-binding protein and named it Taxilin. Taxilin was a protein with a calculated Mr of 61,890 and 546 amino acids. Taxilin had an extraordinarily long coiled-coil region in its C-terminal half. Taxilin bound to syntaxin-1 and -4 as well as syntaxin-3 but not to syntaxin-7 or -8. Taxilin was ubiquitously expressed. A precise function of Taxilin in intracellular vesicle transport is not known. However, since Taxilin affected Ca^<2+>-dependent exocytosis in PC12 cells, it is at least possible that Taxilin is involved in Ca^<2+>-dependent exocytosis in neuroendocrine cells. In future, we attempt to reveal precise functions of α-fodrin and Taxilin in the formation of raft.

越来越多的证据表明，质膜上微结构域的形成是细胞内信号转导的重要过程。特别是，筏的形成对于通过 GPI 锚定蛋白进行细胞内信号转导至关重要。先前已揭示 Syntaxin-3 是 SNARWE 机制的一个组成部分，通过调节细胞内囊泡运输参与筏的形成。然而，syntaxin-3 调控筏形成的分子机制尚不清楚。然后，在本研究中，我们尝试鉴定新的 Syntaxin-3 相互作用分子。我们发现 α-fodrin（亚膜细胞骨架的主要成分）与 Syntaxin-3 相互作用。 α-Fodrin 与syntaxin-1、syntaxin-4 以及syntaxin-3 结合，但不与syntaxin-7 或-8 结合。 α-Fodrin 和 nc18 或 SNAP-25 与 Syntaxin-1 相互作用。这些结果表明 SNARE 复合物的形成受 α-fodrin 调节，并表明质膜下方的肌动蛋白/fodrin 网络参与了筏的形成。此外，我们还发现了一种新的突触融合蛋白结合蛋白，并将其命名为 Taxilin。 Taxilin 是一种蛋白质，经计算，Mr 为 61,890 和 546 个氨基酸。塔西林的 C 末端有一个非常长的卷曲螺旋区域。 Taxilin 与 syntaxin-1 和 -4 以及 syntaxin-3 结合，但不与 syntaxin-7 或 -8 结合。紫杉素广泛表达。 Taxilin 在细胞内囊泡运输中的精确功能尚不清楚。然而，由于Taxilin影响PC12细胞中Ca 2+ 依赖性胞吐作用，因此Taxilin至少有可能参与神经内分泌细胞中Ca 2+ 依赖性胞吐作用。未来，我们试图揭示α-fodrin和Taxilin在筏形成中的精确功能。

项目成果

Kobayashi, N., Mori, Y., Nakano, S., Tsubokou, Y., Shirataki, H., and Matsuoka, H.: "Celiprolol stimulates endothelial nitric oxide synthase expression and improves myocardinal remodeling in deoxycorticosterone acetate-salt hypertensive rats"J. Hypertens.

Kobayashi, N.、Mori, Y.、Nakano, S.、Tsubokou, Y.、Shirataki, H. 和 Matsuoka, H.：“塞利洛尔刺激内皮一氧化氮合酶表达并改善脱氧皮质酮醋酸盐高血压大鼠的心肌重塑

Kobayashi, N., Mori, Y., Nakano, S., Tsubokou, Y., Kobayashi, T., Shirataki, H., and Matsuoka, H.: "TCV-116 stimulates eNOS and caveolin-1 expression and improves coronary microvascular remodeling in normotensive and angiotensin II-induced hypertensive ra

Kobayashi, N.、Mori, Y.、Nakano, S.、Tsubokou, Y.、Kobayashi, T.、Shirataki, H. 和 Matsuoka, H.：“TCV-116 刺激 eNOS 和 Caveolin-1 表达并改善冠状动脉

Nakano M., Nogami S., Sato S., Terano A., Shirataki H.: "Interaction of syntaxin with α-fodrin, a major component of the submembranous cytoskeleton."Biochem. Biophys. Res. Commun.. 288巻・2号. 468-475 (2001)

Nakano M.、Nogami S.、Sato S.、Terano A.、Shirataki H.：“突触蛋白与膜下细胞骨架的主要成分的相互作用。”Biochem.Biophys，第 288 卷。 2. 468-475 (2001)

Nogami S., Satoh S., Nakano M., Shimizu H., Fukushima H., Maruyama A., Terano A., Shirataki H: "Taxilin ; a novel syntaxin-binding protein that is involved in Ca^<2+>-dependent exocytosis in neuroendocrine cells"Genes to Cells. 8巻. 17-28 (2002)

Nogami S.、Satoh S.、Nakano M.、Shimizu H.、Fukushima H.、Maruyama A.、Terano A.、Shirataki H：“Taxilin；一种新型突触结合蛋白，参与 Ca^<2+> -神经内分泌细胞中的依赖性胞吐作用“Genes to Cells.Volume 8. 17-28 (2002)

Ishihara S., Hirata A., Minemura M., Nogami S., Ohya Y.: "A mutation in SPC42, which encodes a component of the spindle pole body, results in production of two-spored asci in Saccharomyces cerevisiae"Mol.Genet.Genomics.. 265巻. 585-595 (2001)

Ishihara S.、Hirata A.、Minemura M.、Nogami S.、Ohya Y.：“编码纺锤体极体成分的 SPC42 中的突变导致酿酒酵母产生双孢子子囊”Mol.基因.基因组学..卷 265. 585-595 (2001)