Hydrogen peroxide depletes phosphatidylinositol-3-phosphate from endosomes in a p38 MAPK-dependent manner and perturbs endocytosis
Hydrogen peroxide depletes phosphatidylinositol-3-phosphate from endosomes in a p38 MAPK-dependent manner and perturbs endocytosis
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DOI：
10.1016/j.bbamcr.2011.01.023
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发表时间：
2011-05-01
影响因子：
5.1
通讯作者：
Murata, Masayuki
中科院分区：
文献类型：
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作者：
Kano, Fumi;Arai, Tamaki;Murata, Masayuki
磷脂酰肌醇-3-磷酸（PI3P）是一种在早期核内体中特异性富集的脂质。考虑到含有PI3P的早期核内体作为一个微结构域来招募含有PI3P结合结构域（FYVE结构域）的蛋白质，PI3P的产生和降解之间的平衡影响了多种过程，包括内吞作用和通过核内体的信号转导。在本文报道的研究中，我们开发了一种新的分析方法，通过将GST-2xFYVE蛋白探针引入半完整细胞中来定量内体中PI3P的数量。GST-2xFYVE探针特异性靶向细胞内含PI3P的核内体，保留其小点状结构，可以半定量测量细胞内PI3P。使用该方法，我们发现用H(2)O(2)处理Hela细胞以p38 mapk依赖的方式降低了内体中PI3P的数量。此外，H(2)O(2)处理延迟了通过各种内吞途径的转运，尤其是早期后的内核体转运；霍乱毒素的逆行转运特别依赖于内体中PI3P的数量。(C) 2011 Elsevier B.V.版权所有
Phosphatidylinositol-3-phosphate (PI3P) is a lipid that is enriched specifically in early endosomes. Given that early endosomes containing PI3P act as a microdomain to recruit proteins that contain a PI3P-binding domain (FYVE domain), the equilibrium between the production and degradation of PI3P influences a variety of processes, including endocytosis and signal transduction via endosomes. In the study reported herein, we have developed a novel analytical method to quantify the amount of PI3P in endosomes by introducing a GST-2xFYVE protein probe into semi-intact cells. The GST-2xFYVE probe was targeted specifically to intracellular PI3P-containing endosomes, which retained their small punctate structure, and allowed the semi-quantitative measurement of intracellular PI3P. Using the method, we found that treatment of Hela cells with H(2)O(2) decreased the amount of PI3P in endosomes in a p38 MAPK-dependent manner. In addition, H(2)O(2) treatment delayed transport through various endocytic pathways, especially post-early endosome transport; the retrograde transport of cholera toxin was especially dependent on the amount of PI3P in endosomes. (C) 2011 Elsevier B.V. All rights reserved.