title: Use of transmembrane FRET to investigate the internalization of glycosylated proteins.
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.
authors.affiliation: 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.
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Haga
authors.forename: Tadashi
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D051275:Glucose Transporter Type 4 / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D018613:Microscopy, Confocal; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D051275:Glucose Transporter Type 4; D006023:Glycoproteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins