GD3 Ganglioside, Expression, Modification and its Biological Function
GD3神经节苷脂、表达、修饰及其生物学功能
基本信息
- 批准号:10480173
- 负责人:
- 金额:$ 2.88万
- 依托单位:
- 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 2000
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The association of ganglioside GD3 with TAG-1, a glycosylphosphatidylinositol-anchored neuronal cell adhesion molecule, was examined by coimmunoprecipitation experiments. Previously, we have shown that the anti-ganglioside GD3 antibody (R24) immunoprecipitated the Src family kinase Lyn from the rat cerebellum, and R24 treatment of primary cerebellar cultures induced Lyn activation and rapid tyrosine phosphorylation of an 80-kDa protein (p80). We now report that R24 coimmunoprecipitates a 135-kDa protein (p135) from primary cerebellar cultures. Treatment with phosphatidylinositol-specitic phospholipase C revealed that p135 was glycosylphosphatidylinositol-anchored to the membrane. It was identified as TAG-1 by sequential immunoprecipitation with an anti-TAG-1 antibody. Antibody-mediated cross-linking of TAG-1 induced Lyn activation and rapid tyrosine phosphorylation of p80. Selective inhibitor for Src family kinases reduced the tyrosine phosphorylation of p80. Sucrose density gradient analysis revealed that the TAG-1 and tyrosine-phosphorylated p80 in cerebellar cultures were present in the lipid raft fraction. These data show that TAG-1 transduces signals via Lyn to p80 in the lipid rafts of the cerebellum. Furthermore, degradation of cell-surface glycosphingolipids by endoglycoceramidase induced an alteration of TAG-1 distribution on an OptiPrep gradient and reduced the TAG-1-mediated Lyn activation and tyrosine phosphorylation of p80. These observations suggest that glycosphingolipids are involved in TAG-1-mediated signaling in lipid rafts.
通过免疫共沉淀实验检查了神经节苷脂 GD3 与 TAG-1(一种糖基磷脂酰肌醇锚定的神经元细胞粘附分子)的关联。之前,我们已经证明抗神经节苷脂GD3抗体(R24)免疫沉淀来自大鼠小脑的Src家族激酶Lyn,并且R24处理原代小脑培养物诱导Lyn激活和80-kDa蛋白(p80)的快速酪氨酸磷酸化。我们现在报道 R24 能够从原代小脑培养物中免疫共沉淀出 135 kDa 的蛋白质 (p135)。用磷脂酰肌醇特异性磷脂酶 C 处理表明 p135 是糖基磷脂酰肌醇锚定在膜上。通过使用抗 TAG-1 抗体进行连续免疫沉淀,将其鉴定为 TAG-1。抗体介导的 TAG-1 交联诱导 Lyn 激活和 p80 的快速酪氨酸磷酸化。 Src 家族激酶的选择性抑制剂可降低 p80 的酪氨酸磷酸化。蔗糖密度梯度分析表明,小脑培养物中的 TAG-1 和酪氨酸磷酸化 p80 存在于脂筏部分中。这些数据表明 TAG-1 通过 Lyn 将信号转导至小脑脂筏中的 p80。此外,神经酰胺内切酶对细胞表面鞘糖脂的降解诱导了 OptiPrep 梯度上 TAG-1 分布的改变,并减少了 TAG-1 介导的 Lyn 激活和 p80 酪氨酸磷酸化。这些观察结果表明,鞘糖脂参与脂筏中 TAG-1 介导的信号传导。
项目成果
期刊论文数量(38)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Kasahara,K.,Watanabe,K.,Takeuchi,K.,Kaneko,H.,Oohira,A.,Yamamoto,T.,Sanai,Y.:: "Involvement of gangliosides in GPI-anchored neuronal cell adhesion molecule TAG-1 signaling in lipid rafts"J.Biol.Chem.. 275. 34701-34709 (2000)
Kasahara,K.,Watanabe,K.,Takeuchi,K.,Kaneko,H.,Oohira,A.,Yamamoto,T.,Sanai,Y.:“神经节苷脂参与 GPI 锚定的神经元细胞粘附分子 TAG-
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Kanai,Y..Kanai-Azuma,M.,Tajima,Y.,Birk,O.S.,Hayashi,Y.,Sanai.Y.: "Identification of a stromal cell type characterized by the secretion of a soluble integrin-binding protein, MFG-E8, in mouse early gonadogenesis."Mech Dev.96, 223-227, (2000). 96. 233-227 (
Kanai,Y..Kanai-Azuma,M.,Tajima,Y.,Birk,O.S.,Hayashi,Y.,Sanai.Y.:“鉴定以分泌可溶性整联蛋白结合蛋白为特征的基质细胞类型,
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Nagase T, Shimoda Y, Sanai Y, Nakamura S, Harii K, Osumi N.: "Differential expression of two glucuronyltransferases synthesizing HNK-1 carbohydrate epitope in the sublineages of the rat myogenic progenitors."Mech Dev.. 98. 145-149 (2000)
Nagase T、Shimoda Y、Sanai Y、Nakamura S、Harii K、Osumi N.:“在大鼠肌原祖细胞亚系中合成 HNK-1 碳水化合物表位的两种葡萄糖醛酸转移酶的差异表达。”Mech Dev.. 98. 145-149
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Shimoda, Y. 他: "Cloning and Expression of a Novel Galactoside β1, 3-Glacronyl-transferase involved in the Biosynthesis of HNK-1 Epitope"J. Biol. Chem.. 274. 17115-17122 (1999)
Shimoda, Y. 等人:“参与 HNK-1 表位生物合成的新型半乳糖苷 β1, 3-Glacronyl-转移酶的克隆和表达”J. Biol. 274. 17115-17122 (1999)
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Kasahara K.,Sanai,Y.: "Functional role of glycosphingolipids in signal transduction via lipid rafts"Glycoconjugate J.. 17. 153-162 (2000)
Kasahara K.,Sanai,Y.:“鞘糖脂在脂筏信号转导中的功能作用”Glycoconjugate J.. 17. 153-162 (2000)
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SANAI Yutaka其他文献
SANAI Yutaka的其他文献
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{{ truncateString('SANAI Yutaka', 18)}}的其他基金
ISOLATION AND CELL BIOLOGICAL CHARACTERIZATION OF SIALYLTRANSFERASES INVOLVED IN GANGLIOSIDE BIOSYNTHESIS
参与神经节苷脂生物合成的唾液酸转移酶的分离和细胞生物学特性
- 批准号:
04670154 - 财政年份:1992
- 资助金额:
$ 2.88万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Isolation and Cell Biological Characterization of Sialyltransferases Involved in Ganglioside Biosynthesis
参与神经节苷脂生物合成的唾液酸转移酶的分离和细胞生物学表征
- 批准号:
02680128 - 财政年份:1990
- 资助金额:
$ 2.88万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Isolation, identification and their regulation of genes responsible for the expression of gangliosides.
负责神经节苷脂表达的基因的分离、鉴定及其调控。
- 批准号:
62580111 - 财政年份:1987
- 资助金额:
$ 2.88万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
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