scholarly journals Arf1/COPI machinery acts directly on lipid droplets and enables their connection to the ER for protein targeting

eLife ◽  
2014 ◽  
Vol 3 ◽  
Author(s):  
Florian Wilfling ◽  
Abdou Rachid Thiam ◽  
Maria-Jesus Olarte ◽  
Jing Wang ◽  
Rainer Beck ◽  
...  
Keyword(s):  
Surface Tension ◽  
Lipid Droplets ◽  
Protein Targeting ◽  
Neutral Lipids ◽  
Vesicle Trafficking ◽  
Metabolic Energy ◽  
Key Enzymes ◽  
Specific Proteins ◽  
Oil Water

Lipid droplets (LDs) are ubiquitous organelles that store neutral lipids, such as triacylglycerol (TG), as reservoirs of metabolic energy and membrane precursors. The Arf1/COPI protein machinery, known for its role in vesicle trafficking, regulates LD morphology, targeting of specific proteins to LDs and lipolysis through unclear mechanisms. Recent evidence shows that Arf1/COPI can bud nano-LDs (∼60 nm diameter) from phospholipid-covered oil/water interfaces in vitro. We show that Arf1/COPI proteins localize to cellular LDs, are sufficient to bud nano-LDs from cellular LDs, and are required for targeting specific TG-synthesis enzymes to LD surfaces. Cells lacking Arf1/COPI function have increased amounts of phospholipids on LDs, resulting in decreased LD surface tension and impairment to form bridges to the ER. Our findings uncover a function for Arf1/COPI proteins at LDs and suggest a model in which Arf1/COPI machinery acts to control ER-LD connections for localization of key enzymes of TG storage and catabolism.

scholarly journals Dual binding motifs underpin the hierarchical association of perilipins1–3 with lipid droplets

2019 ◽  
Vol 30 (5) ◽  
pp. 703-716 ◽  
Author(s):  
Dalila Ajjaji ◽  
Kalthoum Ben M'barek ◽  
Michael L. Mimmack ◽  
Cheryl England ◽  
Haya Herscovitz ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Tissue Type ◽  
Binding Motifs ◽  
Amino Terminal ◽  
A Cell ◽  
Oil Water ◽  
Carboxy Terminal ◽  
Hierarchical Manner

Lipid droplets (LDs) in all eukaryotic cells are coated with at least one of the perilipin (Plin) family of proteins. They all regulate key intracellular lipases but do so to significantly different extents. Where more than one Plin is expressed in a cell, they associate with LDs in a hierarchical manner. In vivo, this means that lipid flux control in a particular cell or tissue type is heavily influenced by the specific Plins present on its LDs. Despite their early discovery, exactly how Plins target LDs and why they displace each other in a “hierarchical” manner remains unclear. They all share an amino-terminal 11-mer repeat (11mr) amphipathic region suggested to be involved in LD targeting. Here, we show that, in vivo, this domain functions as a primary highly reversible LD targeting motif in Plin1–3, and, in vitro, we document reversible and competitive binding between a wild-type purified Plin1 11mr peptide and a mutant with reduced binding affinity to both “naked” and phospholipid-coated oil–water interfaces. We also present data suggesting that a second carboxy-terminal 4-helix bundle domain stabilizes LD binding in Plin1 more effectively than in Plin2, whereas it weakens binding in Plin3. These findings suggest that dual amphipathic helical regions mediate LD targeting and underpin the hierarchical binding of Plin1–3 to LDs.

scholarly journals YPR139c/LOA1encodes a novel lysophosphatidic acid acyltransferase associated with lipid droplets and involved in TAG homeostasis

2012 ◽  
Vol 23 (2) ◽  
pp. 233-246 ◽  
Author(s):  
Sophie Ayciriex ◽  
Marina Le Guédard ◽  
Nadine Camougrand ◽  
Gisèle Velours ◽  
Mario Schoene ◽  
...  
Keyword(s):  
Lysophosphatidic Acid ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
In Vitro Assays ◽  
Metabolic Labeling ◽  
Lysophosphatidic Acid Acyltransferase ◽  
Escherichia Coli Cells ◽  
Type Strains

For many years, lipid droplets (LDs) were considered to be an inert store of lipids. However, recent data showed that LDs are dynamic organelles playing an important role in storage and mobilization of neutral lipids. In this paper, we report the characterization of LOA1 (alias VPS66, alias YPR139c), a yeast member of the glycerolipid acyltransferase family. LOA1 mutants show abnormalities in LD morphology. As previously reported, cells lacking LOA1 contain more LDs. Conversely, we showed that overexpression results in fewer LDs. We then compared the lipidome of loa1Δ mutant and wild-type strains. Steady-state metabolic labeling of loa1Δ revealed a significant reduction in triacylglycerol content, while phospholipid (PL) composition remained unchanged. Interestingly, lipidomic analysis indicates that both PLs and glycerolipids are qualitatively affected by the mutation, suggesting that Loa1p is a lysophosphatidic acid acyltransferase (LPA AT) with a preference for oleoyl-CoA. This hypothesis was tested by in vitro assays using both membranes of Escherichia coli cells expressing LOA1 and purified proteins as enzyme sources. Our results from purification of subcellular compartments and proteomic studies show that Loa1p is associated with LD and active in this compartment. Loa1p is therefore a novel LPA AT and plays a role in LD formation.

scholarly journals A Unique Junctional Interface at Contact Sites Between the Endoplasmic Reticulum and Lipid Droplets

2021 ◽  
Vol 9 ◽  
Author(s):  
Vineet Choudhary ◽  
Roger Schneiter
Keyword(s):  
Endoplasmic Reticulum ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Close Contact ◽  
Lipid Storage ◽  
Metabolic Energy ◽  
Lipid Monolayer ◽  
Storage Diseases ◽  
Contact Sites ◽  
Ordered Assembly

Lipid droplets (LDs) constitute compartments dedicated to the storage of metabolic energy in the form of neutral lipids. LDs originate from the endoplasmic reticulum (ER) with which they maintain close contact throughout their life cycle. These ER–LD junctions facilitate the exchange of both proteins and lipids between these two compartments. In recent years, proteins that are important for the proper formation of LDs and localize to ER–LD junctions have been identified. This junction is unique as it is generally believed to invoke a transition from the ER bilayer membrane to a lipid monolayer that delineates LDs. Proper formation of this junction requires the ordered assembly of proteins and lipids at specialized ER subdomains. Without such a well-ordered assembly of LD biogenesis factors, neutral lipids are synthesized throughout the ER membrane, resulting in the formation of aberrant LDs. Such ectopically formed LDs impact ER and lipid homeostasis, resulting in different types of lipid storage diseases. In response to starvation, the ER–LD junction recruits factors that tether the vacuole to these junctions to facilitate LD degradation. In addition, LDs maintain close contacts with peroxisomes and mitochondria for metabolic channeling of the released fatty acids toward beta-oxidation. In this review, we discuss the function of different components that ensure proper functioning of LD contact sites, their role in lipogenesis and lipolysis, and their relation to lipid storage diseases.

scholarly journals Lecithin:Retinol Acyl Transferase (LRAT) induces the formation of lipid droplets

2019 ◽  
Author(s):  
Martijn R. Molenaar ◽  
Tsjerk A. Wassenaar ◽  
Kamlesh K. Yadav ◽  
Alexandre Toulmay ◽  
Muriel C. Mari ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Yeast Cells ◽  
Acyl Transferase ◽  
Hydrophobic Core ◽  
Retinyl Ester ◽  
Steryl Esters ◽  
Retinyl Esters

AbstractLipid droplets are unique and nearly ubiquitous organelles that store neutral lipids in a hydrophobic core, surrounded by a monolayer of phospholipids. The primary neutral lipids are triacylglycerols and steryl esters. It is not known whether other classes of neutral lipids can form lipid droplets by themselves. Here we show that production of retinyl esters by lecithin:retinol acyl transferase (LRAT) in yeast cells, incapable of producing triacylglycerols and steryl esters, causes the formation of lipid droplets. By electron microscopy, these lipid droplets are morphologically indistinguishable from those in wild-type cells. In silico and in vitro experiments confirmed the propensity of retinyl esters to segregate from membranes and to form lipid droplets. The hydrophobic N-terminus of LRAT displays preferential interactions with retinyl esters in membranes and promotes the formation of large retinyl ester-containing lipid droplets in mammalian cells. Our combined data indicate that the molecular design of LRAT is optimally suited to allow the formation of characteristic large lipid droplets in retinyl ester-storing cells.

Intracellular communication between lipid droplets and peroxisomes: the Janus face of PEX19

2018 ◽  
Vol 399 (7) ◽  
pp. 741-749 ◽  
Author(s):  
Bianca Schrul ◽  
Wolfgang Schliebs
Keyword(s):  
Metabolic Pathways ◽  
Lipid Droplets ◽  
Environmental Changes ◽  
Neutral Lipids ◽  
Nutrient Supply ◽  
Metabolic Energy ◽  
Central Question ◽  
And Function ◽  
Metabolic Output

Abstract In order to adapt to environmental changes, such as nutrient availability, cells have to orchestrate multiple metabolic pathways, which are catalyzed in distinct specialized organelles. Lipid droplets (LDs) and peroxisomes are both endoplasmic reticulum (ER)-derived organelles that fulfill complementary functions in lipid metabolism: Upon nutrient supply, LDs store metabolic energy in the form of neutral lipids and, when energy is needed, supply fatty acids for oxidation in peroxisomes and mitochondria. How these organelles communicate with each other for a concerted metabolic output remains a central question. Here, we summarize recent insights into the biogenesis and function of LDs and peroxisomes with emphasis on the role of PEX19 in these processes.

scholarly journals The effect of methyl-lidocaine on the biosynthesis of phospholipids de novo in the isolated hamster heart

1992 ◽  
Vol 285 (1) ◽  
pp. 161-166 ◽  
Author(s):  
P G Tardi ◽  
R Y K Man ◽  
P C Choy
Keyword(s):  
De Novo ◽  
Neutral Lipids ◽  
Phosphatidate Phosphatase ◽  
Perfused Heart ◽  
Intracellular Pool ◽  
Key Enzymes ◽  
Acidic Phospholipids ◽  
Hamster Heart ◽  
Pool Sizes

Methyl-lidocaine is an amphiphilic agent which has been used as an experimental anti-arrhythmic drug. When hamster hearts were perfused with labelled glycerol, the presence of methyl-lidocaine in the perfusate was found to enhance the labelling in phosphatidylserine, phosphatidylinositol, diacylglycerol and triacylglycerol. However, the labelling of phosphatidylcholine and phosphatidylethanolamine was not significantly changed by methyl-lidocaine treatment. Assays in vitro for the enzymes involved in the synthesis of neutral lipids and acidic phospholipids revealed that phosphatidate phosphatase and CTP: phosphatidate cytidylyltransferase activities were stimulated by methyl-lidocaine. The intracellular pool sizes of diacylglycerol and CDP-diacylglycerol were also elevated. We postulate that the enhanced syntheses of the neutral lipids and acidic phospholipids in the methyl-lidocaine-perfused heart were mediated via the direct activation of the key enzymes in the biosynthesis of these lipids de novo.

scholarly journals Adiposome Targeting and Enzymatic Activity of Lipid Droplet-Specific Proteins

2020 ◽  
Author(s):  
Xuejing Ma ◽  
Zelun Zhi ◽  
Shuyan Zhang ◽  
Chang Zhou ◽  
Adam Mechler ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Lipid Droplet ◽  
Protein Targeting ◽  
Phosphorylation Site ◽  
Specific Protein ◽  
Scatchard Analysis ◽  
Specific Proteins ◽  
Perilipin 2

SUMMARYNew strategies to decode the specific protein targeting mechanism on lipid droplet (LD) are urgently needed. Using adiposome, the LD binding of perilipin 2 (PLIN2), perilipin 3 (PLIN3), and adipose triglyceride lipase (ATGL) were studied. Scatchard analysis found that the binding of PLIN2 to the adiposome surface was saturable, pointing to a specific membrane binding partner. Phosphatidylinositol (PI) was found to inhibit PLIN2 binding while it did not impede PLIN3. Structural analysis combined with mutagenesis revealed that the 73rd glutamic acid of PLIN2 is significant for the effect of PI on the protein binding. The presence of PI significantly stimulated the activity of ATGL in vitro. The phosphorylation site mutants of ATGL were found reducing the lipase activity in the adiposome system. Our study demonstrates the utility of adiposome as a powerful, manipulatable model system for the characterization of LD binding and enzymatic activity of LD proteins in vitro.

scholarly journals Universal phase behaviors of intracellular lipid droplets

2019 ◽  
Vol 116 (51) ◽  
pp. 25440-25445 ◽  
Author(s):  
Shunsuke F. Shimobayashi ◽  
Yuki Ohsaki
Keyword(s):  
Phase Transition ◽  
Phase Diagram ◽  
Lipid Droplets ◽  
Energy Homeostasis ◽  
Neutral Lipids ◽  
Droplet Surface ◽  
Molecular Organization ◽  
Cholesteryl Esters

Lipid droplets are cytoplasmic microscale organelles involved in energy homeostasis and handling of cellular lipids and proteins. The core structure is mainly composed of two kinds of neutral lipids, triglycerides and cholesteryl esters, which are coated by a phospholipid monolayer and proteins. Despite the liquid crystalline nature of cholesteryl esters, the connection between the lipid composition and physical states is poorly understood. Here, we present a universal intracellular phase diagram of lipid droplets, semiquantitatively consistent with the in vitro phase diagram, and reveal that cholesterol esters cause the liquid–liquid crystal phase transition under near-physiological conditions. We moreover combine in vivo and in vitro studies, together with the theory of confined liquid crystals, to suggest that the radial molecular alignments in the liquid crystallized lipid droplets are caused by an anchoring force at the droplet surface. Our findings on the phase transition of lipid droplets and resulting molecular organization contribute to a better understanding of their biological functions and diseases.

scholarly journals Universal phase behaviors of intracellular lipid droplets

2019 ◽  
Author(s):  
Shunsuke F. Shimobayashi ◽  
Yuki Ohsaki
Keyword(s):  
Phase Transition ◽  
Phase Diagram ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Droplet Surface ◽  
Molecular Organization ◽  
Cholesteryl Esters ◽  
Intracellular Lipid

Lipid droplets are cytoplasmic micro-scale organelles involved in energy homeostasis and handling of cellular lipids and proteins. The core structure is mainly composed of two kinds of neutral lipids, triglycerides and cholesteryl esters, which are coated by a phospholipid monolayer and proteins. Despite the liquid crystalline nature of cholesteryl esters, the connection between the lipid composition and physical states is poorly understood. Here, we present the first universal intracellular phase diagram of lipid droplets, semi-quantitatively consistent with the in vitro phase diagram, and reveal that cholesterol esters cause the liquid-liquid crystal phase transition under near-physiological conditions. The internal molecules of the liquid crystallized lipid droplets are aligned radially. We moreover combine in vivo and in vitro studies, together with the theory of confined liquid crystals, to suggest that the radial molecular alignments in intracellular lipid droplets are caused by an anchoring force at the droplet surface. Our findings on the phase transition of lipid droplets and resulting molecular organization contribute to a better understanding of their biological functions and diseases.

scholarly journals Retinyl esters form lipid droplets independently of triacylglycerol and seipin

2021 ◽  
Vol 220 (10) ◽  
Author(s):  
Martijn R. Molenaar ◽  
Kamlesh K. Yadav ◽  
Alexandre Toulmay ◽  
Tsjerk A. Wassenaar ◽  
Muriel C. Mari ◽  
...  
Keyword(s):  
Lipid Bilayers ◽  
Lipid Droplet ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Yeast Cells ◽  
In Vitro Experiments ◽  
Steryl Esters ◽  
Retinyl Esters

Lipid droplets store neutral lipids, primarily triacylglycerol and steryl esters. Seipin plays a role in lipid droplet biogenesis and is thought to determine the site of lipid droplet biogenesis and the size of newly formed lipid droplets. Here we show a seipin-independent pathway of lipid droplet biogenesis. In silico and in vitro experiments reveal that retinyl esters have the intrinsic propensity to sequester and nucleate in lipid bilayers. Production of retinyl esters in mammalian and yeast cells that do not normally produce retinyl esters causes the formation of lipid droplets, even in a yeast strain that produces only retinyl esters and no other neutral lipids. Seipin does not determine the size or biogenesis site of lipid droplets composed of only retinyl esters or steryl esters. These findings indicate that the role of seipin in lipid droplet biogenesis depends on the type of neutral lipid stored in forming droplets.

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