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

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 t...

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Bibliographic Details
Published in:Biochemical journal 2001-09, Vol.358 (Pt 2), p.517-522
Main Authors: Lisinski, I, Schürmann, A, Joost, H G, Cushman, S W, Al-Hasani, H
Format: Article
Language:English
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
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Summary: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.
ISSN:0264-6021
1470-8728
DOI:10.1042/0264-6021:3580517