Fluorogenic probes reveal a role of GLUT4 N-glycosylation in intracellular trafficking

Shinya Hirayama, Yuichiro Hori, Zsolt Benedek, Tadashi Suzuki, Kazuya Kikuchi

Research output: Contribution to journalArticlepeer-review

40 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)853-859
Number of pages7
JournalNature Chemical Biology
Volume12
Issue number10
DOIs
Publication statusPublished - Oct 1 2016
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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