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Analysis of amino and carboxy terminal GLUT-4 targeting motifs in 3T3-L1 adipocytes using an endosomal ablation technique.

Abstract：

:The targeting of the insulin-responsive glucose transporter, GLUT-4, to an intracellular compartment in adipocytes and muscle is one of the key features responsible for the unique insulin sensitivity of this transporter. Through expression of epitope-tagged GLUT-4 mutants in 3T3-L1 adipocytes, two motifs have been identified as playing a central role in GLUT-4 targeting: FQQI in the amino terminus and a di-leucine motif in the carboxy terminus. The goal of this study was to explore the role of these targeting motifs in the intracellular sorting of GLUT-4 using the Tf-HRP ablation technique. This technique provides a quantitative assessment of the amount of GLUT-4 located in recycling endosomes. In basal adipocytes, we find that approximately 40% of GLUT-4 is ablated following Tf-HRP loading. In contrast, here we demonstrate that the intracellular pool of a mutant in which F5 was mutated to A5 is localized to the recycling endosomal pathway, suggesting that the amino terminal FQQI motif functions in trafficking GLUT-4 from early endosomes. In contrast, GLUT-4 in which L489L490 was mutated to A489A490 was localized predominantly to a nonablated compartment. These data imply a role for the di-leucine motif in sorting from a separate intracellular compartment, such as the TGN. Our findings are discussed within the context of a revised multicompartment model for GLUT-4 trafficking in adipocytes, in which mutations in either the FQQI or LL motifs result in the altered subcellular trafficking of GLUT-4 between multiple intracellular compartments.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Melvin DR,Marsh BJ,Walmsley AR,James DE,Gould GW

doi

10.1021/bi980988y

subject

Has Abstract

pub_date

1999-02-02 00:00:00

pages

1456-62

issue

5

eissn

0006-2960

issn

1520-4995

pii

bi980988y

journal_volume

38

pub_type

杂志文章

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