title: Hydrophobic sequences target and anchor perilipin A to lipid droplets.
abstract: Perilipins regulate triacylglycerol storage and hydrolysis in adipocytes. The central 25% of the perilipin A sequence, including three hydrophobic sequences (H1, H2, and H3) and an acidic region, targets and anchors perilipins to lipid droplets. Thus, we hypothesized that H1, H2, and H3 are targeting and anchoring motifs. We now show that deletion of any single hydrophobic sequence or combinations of H1 and H3 or H2 and H3 does not prevent targeting of the mutated perilipin to lipid droplets. In contrast, mutated perilipin lacking H1 and H2 showed reduced targeting, whereas perilipin lacking H1, H2, and H3 targeted poorly to lipid droplets; thus, H3 is a weak targeting signal and either H1 or H2 is required for optimal targeting. Complete elimination of perilipin targeting was observed only when all three hydrophobic sequences were deleted in combination with either the acidic region or N-terminal sequences predicted to form amphipathic beta-strands. Unlike intact perilipin A, mutated perilipin lacking either H1 and H2 or H1, H2, and H3 was released from lipid droplets after alkaline carbonate treatment, suggesting that these forms are loosely associated with lipid droplets. The three hydrophobic sequences play a major role in targeting and anchoring perilipins to lipid droplets.
authors.affiliation: Department of Nutritional Sciences, Rutgers, The State University of New Jersey, New Brunswick, NJ 08901, USA.
authors.forename: Vidya
authors.initials: V
authors.lastname: Subramanian
authors.forename: Anne
authors.initials: A
authors.lastname: Garcia
authors.forename: Anna
authors.initials: A
authors.lastname: Sekowski
authors.forename: Dawn L
authors.initials: DL
authors.lastname: Brasaemle
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000378:metabolism; D020816:Amino Acid Motifs; D000818:Animals; D015152:Blotting, Northern; D002254:Carbonates / Q000494:pharmacology; D002352:Carrier Proteins; D005347:Fibroblasts / Q000378:metabolism; D015151:Immunoblotting; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D009154:Mutation; D000070778:Perilipin-1; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism
chemical_list: D002254:Carbonates; D002352:Carrier Proteins; D008055:Lipids; D000070778:Perilipin-1; D010750:Phosphoproteins; D012333:RNA, Messenger; D014280:Triglycerides