Study on the topology of the GPI anchor assembly

GPI锚固组件的拓扑研究

• 批准号: 09680707
• 负责人: HIROSE Shinichi
• 金额: $ 1.86万
• 依托单位: Fukuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680707/
• 关键词: GPI anchor

The glycophosphatidylinositol (GPI) anchor is preassembled and transferred to the c-terminus of the corresponding nascent proteins in the ER. The assembly starts in the luminal side of the ER and proceeds between the cytosolic and luminal sides of the ER. Thus, precursors of GPI anchor have to flip-flop between the lipid bilayer during the assembly. The mechanisms underlying this flip-flop has not been well characterized, nor identified are cues for the flip-flop. The GPI anchor proteins on some cell lines are known to be acylated while most of the GPI proteins are not acylated. This implied that acylation involved in the flip-flop as a cue. We have investigated the timing of the flip-flop in the relation, with the acylation of the precursors on erythroleukemia cells. The GPI anchor including the precursors of which inositol residue is acylated are resistant GPI specific pshopholipase C digestion.The deacylation took after the transfer of the preassembled GPI to the target nascent proteins. This findings thus suggested that the acylation is not involved in the flip-flop.

糖磷脂酰肌醇（GPI）锚被预先组装并转移到ER中相应新生蛋白的C-末端。组装开始于ER的腔侧，并在ER的胞质侧和腔侧之间进行。因此，GPI锚的前体在组装期间必须在脂质双层之间翻转。这种触发器的机制尚未得到很好的表征，也没有确定的触发器的线索。已知一些细胞系上的GPI锚蛋白是酰化的，而大多数GPI蛋白是不酰化的。这意味着酰化作为一种线索参与了这种翻转。我们已经调查了触发器的时间与红白血病细胞上前体的酰化的关系。GPI锚包括其肌醇残基被酰化的前体，其抗GPI特异性的磷脂酶C消化。因此，这一发现表明酰化不参与触发。

项目成果

Tujioka, H., et al.: "Posttranslational Modification of Glycosylphosphatidylinositol (GPI)-Specific Phospholipase D and Its Activity in Cleavage of GPI Anchors"Biochemical and Biophysical Research Communications. 251. 737-743 (1998)

Tujioka, H. 等人：“糖基磷脂酰肌醇 (GPI) 特异性磷脂酶 D 的翻译后修饰及其在 GPI 锚定裂解中的活性”生物化学和生物物理研究通讯。

Tsujioka H,et al.: "Intracellular cleavage of glycosylphosphatidylinositol by phospholipase D induces activation of protein kinase Calpha"Biochem J. 342(Pt2). 449-555 (1999)

Tsujioka H 等人：“磷脂酶 D 对糖基磷脂酰肌醇的细胞内裂解诱导蛋白激酶 Cα 的激活”Biochem J. 342（Pt2）。

M.Sohda,Y Misumi,A Yano et al.: "Phosphorylation of vesicle doking proteins p115 regulatesits association with the Goldi membrane" Journal of Biological Chemistry. 273(in press).

M.Sohda、Y Misumi、A Yano 等人：“囊泡 doking 蛋白 p115 的磷酸化调节其与 Goldi 膜的关联”《生物化学杂志》。

Chen, R., et al.: "Mammalian glycophosphatidylinositol anchor transfer to proteins and posttransfer deacylation"Proc. Natl. Acad. Sci.. 95. 9512-9517 (1998)

Chen, R. 等人：“哺乳动物糖磷脂酰肌醇锚定转移到蛋白质和转移后脱酰化”Proc。

Sohda, M., et al.: "Phosphorylation of the Vesicle Docking Protein p115 Regulates Its Association with the Golgi MembraneィイD1*ィエD1"The Journal of Biological Chemistry. 273. 5385-5388 (1998)

Sohda, M., 等人：“囊泡对接蛋白 p115 的磷酸化调节其与高尔基膜 D1*D1 的关联”《生物化学杂志》273. 5385-5388 (1998)。