Induced translocation of glycosylphosphatidylinositol-anchored proteins from lipid droplets to adiposomes in rat adipocytes.
OBJECTIVE
Adipocytes release membrane vesicles called adiposomes, which harbor the glycosylphosphatidylinositol-anchored proteins (GPI proteins), Gce1 and CD73, after induction with palmitate, H(2)O(2) and the sulphonylurea drug glimepiride. The role of lipid droplets (LD) in trafficking of GPI proteins from detergent-insoluble, glycolipid-enriched, plasma membrane microdomains (DIGs) to adiposomes in rat adipocytes was studied.


METHODS
Redistribution of Gce1 and CD73 was followed by pulse-chase and long-term labelling, western blot analysis and activity determinations with subcellular fractions and cell-free systems exposed to palmitate, H(2)O(2) and glimepiride.


RESULTS
In response to these signals, Gce1 and CD73 disappeared from DIGs, then transiently appeared in LD and finally were released into adiposomes from small, and, more efficiently, large adipocytes. From DIGs to LD, Gce1 and CD73 were accompanied by cholesterol. Cholesterol depletion from DIGs or LD caused accumulation at DIGs or accelerated loss from LD and release into adiposomes, respectively, of the GPI proteins. Blockade of translocation of Gce1, CD73, caveolin-1 and perilipin-A from DIGs to LD blocked LD biogenesis and long term-accumulation of LD interfered with induced release of the GPI proteins into adiposomes. GPI protein release was up-regulated upon long term-depletion of LD. Adiposomes were released by a DIGs-based cell-free system, but only in presence of LD.


CONCLUSIONS
GPI proteins are translocated from DIGs to LD prior to their release into adiposomes, which is regulated by cholesterol, LD content and LD biogenesis. This detour may serve to transfer information about the LD content and to control lipolysis/esterification between large and small adipocytes via GPI protein-harbouring adiposomes.
Müller
G
G
Sanofi-Aventis Pharma, R & D, Therapeutic Department Metabolism, Frankfurt am Main, Germany. Guenter.Mueller@sanofi-aventis.com
Jung
C
C
Wied
S
S
Biemer-Daub
G
G
D015720:5'-Nucleotidase / Q000378:metabolism*; D017667:Adipocytes / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D002352:Carrier Proteins; D051242:Caveolin 1 / Q000378:metabolism; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D055354:Exosomes / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism*; D010168:Palmitates / Q000494:pharmacology; D000070778:Perilipin-1; D010750:Phosphoproteins / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism*; D021381:Protein Transport; D051381:Rats; D017207:Rats, Sprague-Dawley; D013453:Sulfonylurea Compounds / Q000494:pharmacology
D002352:Carrier Proteins; C495537:Cav1 protein, rat; D051242:Caveolin 1; D017261:Glycosylphosphatidylinositols; D010168:Palmitates; D000070778:Perilipin-1; D010750:Phosphoproteins; D013453:Sulfonylurea Compounds; C057619:glimepiride; D002784:Cholesterol; D006861:Hydrogen Peroxide; D015720:5'-Nucleotidase; D010727:Phosphoric Diester Hydrolases
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