利用跨膜荧光共振能量转移技术研究糖基化蛋白的内化作用。

Use of transmembrane FRET to investigate the internalization of glycosylated proteins.

作者信息

Haga Yoshimi, Suzuki Tadashi

机构信息

Glycometabolome Team, Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center for Systems Chemical Biology, RIKEN Global Research Cluster, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.

出版信息

Methods Mol Biol. 2014;1174:225-30. doi: 10.1007/978-1-4939-0944-5_15.

DOI:10.1007/978-1-4939-0944-5_15

PMID:24947385

Abstract

The importance of glycans in various cellular events, especially intracellular and intercellular trafficking of proteins, has been reported in numerous studies. Here, we present a novel method to monitor endocytosis of proteins of interest bearing a specific glycan modification. Using a fluorescence resonance energy transfer technique, we investigated the role of glycan structure on the internalization of insulin-responsive glucose transporter GLUT4. We found that sialylated glycoforms of GFP-tagged GLUT4 appear to be internalized more slowly than non-sialylated GLUT4 upon insulin removal. This novel glycan imaging tool allows probing functional roles of specific glycan modifications in endocytosis of various proteins.

摘要

许多研究报道了聚糖在各种细胞事件中的重要性，尤其是蛋白质的细胞内和细胞间运输。在此，我们提出了一种新方法，用于监测带有特定聚糖修饰的目标蛋白质的内吞作用。利用荧光共振能量转移技术，我们研究了聚糖结构对胰岛素反应性葡萄糖转运蛋白GLUT4内化的作用。我们发现，在去除胰岛素后，绿色荧光蛋白标记的GLUT4的唾液酸化糖型似乎比非唾液酸化的GLUT4内化得更慢。这种新型聚糖成像工具能够探究特定聚糖修饰在各种蛋白质内吞作用中的功能作用。