title: Proteomic analysis of lipid droplets from Caco-2/TC7 enterocytes identifies novel modulators of lipid secretion.
abstract: In enterocytes, the dynamic accumulation and depletion of triacylglycerol (TAG) in lipid droplets (LD) during fat absorption suggests that cytosolic LD-associated TAG contribute to TAG-rich lipoprotein (TRL) production. To get insight into the mechanisms controlling the storage/secretion balance of TAG, we used as a tool hepatitis C virus core protein, which localizes onto LDs, and thus may modify their protein coat and decrease TRL secretion. We compared the proteome of LD fractions isolated from Caco-2/TC7 enterocytes expressing or not hepatitis C virus core protein by a differential proteomic approach (isobaric tag for relative and absolute quantitation (iTRAQ) labeling coupled with liquid chromatography and tandem mass spectrometry). We identified 42 proteins, 21 being involved in lipid metabolism. Perilipin-2/ADRP, which is suggested to stabilize long term-stored TAG, was enriched in LD fractions isolated from Caco-2/TC7 expressing core protein while perilipin-3/TIP47, which is involved in LD synthesis from newly synthesized TAG, was decreased. Endoplasmic reticulum-associated proteins were strongly decreased, suggesting reduced interactions between LD and endoplasmic reticulum, where TRL assembly occurs. For the first time, we show that 17β-hydroxysteroid dehydrogenase 2 (DHB2), which catalyzes the conversion of 17-keto to 17 β-hydroxysteroids and which was the most highly enriched protein in core expressing cells, is localized to LD and interferes with TAG secretion, probably through its capacity to inactivate testosterone. Overall, we identified potential new players of lipid droplet dynamics, which may be involved in the balance between lipid storage and secretion, and may be altered in enterocytes in pathological conditions such as insulin resistance, type II diabetes and obesity.
authors.affiliation: Université Pierre et Marie Curie, UMR S 872, Les Cordeliers, Paris, France.
authors.forename: Frauke
authors.initials: F
authors.lastname: Beilstein
authors.forename: Julien
authors.initials: J
authors.lastname: Bouchoux
authors.forename: Monique
authors.initials: M
authors.lastname: Rousset
authors.forename: Sylvie
authors.initials: S
authors.lastname: Demignot
mesh_terms: D018938:Caco-2 Cells; D002853:Chromatography, Liquid / Q000379:methods; D003600:Cytosol / Q000378:metabolism; D017931:DNA Primers; D020895:Enterocytes / Q000166:cytology*; D004960:Estradiol Dehydrogenases / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016174:Hepacivirus; D006801:Humans; D050356:Lipid Metabolism; D008856:Microscopy, Fluorescence; D010957:Plasmids; D040901:Proteomics / Q000379:methods*; D013347:Subcellular Fractions; D053719:Tandem Mass Spectrometry / Q000379:methods; D014280:Triglycerides / Q000378:metabolism; D014758:Viral Core Proteins / Q000378:metabolism
chemical_list: D017931:DNA Primers; D014280:Triglycerides; D014758:Viral Core Proteins; D049452:Green Fluorescent Proteins; D004960:Estradiol Dehydrogenases; C523763:HSD17B2 protein, human