Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking
Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking
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DOI：
10.1038/nchembio.2156
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发表时间：
2016-10-01
影响因子：
14.8
通讯作者：
Kikuchi, Kazuya
中科院分区：
文献类型：
--
作者：
Hirayama, Shinya;Hori, Yuichiro;Kikuchi, Kazuya
葡萄糖转运蛋白4 （GLUT4）是一种n -糖基化蛋白，通过调节蛋白质易位维持葡萄糖稳态。迄今为止，尚不清楚GLUT4的n -聚糖是否参与其细胞内运输。在这里，为了阐明n -聚糖的作用，我们开发了荧光探针，标记细胞质和质膜蛋白，用于GLUT4易位的多色成像。其中一种探针是细胞无关的，选择性地检测胞外化的GLUT4。使用该探针，我们验证了运输的“日志”，其中n-聚糖缺陷的GLUT4在胰岛素刺激下瞬间易位到细胞膜上，并迅速内化而不保留在细胞膜上。结果强烈提示n -聚糖在细胞膜上的GLUT4保留中起作用。本研究显示了荧光探针的实用性，并表明该成像工具将适用于研究各种具有动态定位变化的膜蛋白。
Glucose transporter 4 (GLUT4) is an N-glycosylated protein that maintains glucose homeostasis by regulating the protein translocation. To date, it has been unclear whether the N-glycan of GLUT4 contributes to its intracellular trafficking. Here, to clarify the role of the N-glycan, we developed fluorogenic probes that label cytoplasmic and plasma-membrane proteins for multicolor imaging of GLUT4 translocation. One of the probes, which is cell impermeant, selectively detected exocytosed GLUT4. Using this probe, we verified the 'log' of the trafficking, in which N-glycan-deficient GLUT4 was transiently translocated to the cell membrane upon insulin stimulation and was rapidly internalized without retention on the cell membrane. The results strongly suggest that the N-glycan functions in the retention of GLUT4 on the cell membrane. This study showed the utility of the fluorogenic probes and indicated that this imaging tool will be applicable for research on various membrane proteins that show dynamic changes in localization.