Targeting of GLUT6 (formerly GLUT9) and GLUT8 in rat adipose cells

• Published in: Biochemical journal Vol. 358; no. Pt 2; pp. 517 - 522
• Main Authors: Lisinski, I, Schürmann, A, Joost, H G, Cushman, S W, Al-Hasani, H
• Format: Journal Article
• Language: English
• Published: England 01.09.2001
• Subjects: Adipocytes - metabolism; Animals; Cell Membrane - metabolism; Cells, Cultured; COS Cells; Dynamins; Glucose Transport Proteins, Facilitative; Glucose Transporter Type 4; GTP Phosphohydrolases - genetics; GTP Phosphohydrolases - metabolism; Hemagglutinins - genetics; Insulin - pharmacology; Intracellular Membranes - metabolism; Male; Monosaccharide Transport Proteins - chemistry; Monosaccharide Transport Proteins - genetics; Monosaccharide Transport Proteins - metabolism; Muscle Proteins; Mutation; Protein Structure, Tertiary; Protein Transport; Rats; Recombinant Fusion Proteins - metabolism; Transfection
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/0264-6021:3580517

The subcellular targeting of the two recently cloned novel mammalian glucose transporters, GLUT6 [previously referred to as GLUT9 [Doege, Bocianski, Joost and Schürmann (2000) Biochem. J. 350, 771-776] and GLUT8, was analysed by expression of haemagglutinin (HA)-epitope-tagged GLUTs in transiently transfected primary rat adipose cells. Similar to HA-GLUT4, both transporters, HA-GLUT6 and HA-GLUT8, were retained in intracellular compartments in non-stimulated cells. In contrast, mutation of the N-terminal dileucine motifs in both constructs led to constitutive expression of the proteins on the plasma membrane. Likewise, when endocytosis was blocked by co-expression of a dominant-negative mutant of the dynamin GTPase, wild-type HA-GLUT6 and HA-GLUT8 accumulated on the cell surface. However, in contrast with HA-GLUT4, no translocation of HA-GLUT6 and HA-GLUT8 to the plasma membrane was observed when the cells were stimulated with insulin, phorbol ester or hyperosmolarity. Thus GLUT6 and GLUT8 appear to recycle in a dynamin-dependent manner between internal membranes and the plasma membrane in rat adipose cells, but are unresponsive to stimuli that induce translocation of GLUT4.