Purification and Characterization of Recombinant Lipid Storage Protein-2 from Drosophila melanogaster

Arrese, Estela L.; Rivera, Laticia; Hamada, Masakazu; Soulages, José L.

doi:10.2174/092986608785849191

Cited by 21 publications

(16 citation statements)

References 25 publications

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“…The identity of this band was confirmed by mass spectrometry. A similar situation was reported for recombinant DmPLIN2 (41.1 kDa) whose observed electrophoretic mobility corresponded to 55 kDa (Arrese et al, 2008b). …”

Section: Resultssupporting

confidence: 83%

“…Recombinant MsPLIN2 was produced in E. coli as a fusion protein with thioredoxin (Trx), Trx-[His]6-Stag-PLIN2 (49.4 kDa) as we previously described for Drosophila PLIN2 (DmPLIN2) (Arrese et al, 2008b). The fusion protein contain a cleavage site for removal of the Stag-PLIN2 portion.…”

Section: Resultsmentioning

confidence: 99%

“…Drosophila PLIN2 is mostly expressed in tissues directly engaged in lipid metabolism, such as larval fat body and ovary, but it was also found to play a role in lipid accumulation in wing imaginal discs (Fauny et al, 2005). Drosophila PLIN2 protein was found to be highly helical protein, soluble in aqueous medium but also able to tightly bind and form stable complexes with phospholipid (Arrese et al, 2008b). Two putative amphipathic helices could mediate lipid binding.…”

Section: Introductionmentioning

confidence: 99%

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Clues on the function of Manduca sexta perilipin 2 inferred from developmental and nutrition-dependent changes in its expression

Chen

Firdaus

Howard

et al. 2017

Insect Biochemistry and Molecular Biology

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Cellular triglycerides (TG) are stored in cytosolic lipid droplets (LDs). Perilipins (PLIN) are a group of LD-proteins that play important roles in the assembly and transport of LDs and in TG metabolism. Two members of the PLIN family are found in insects (PLIN1 & 2 or Lsd1 & 2). We have cloned and expressed Manduca sexta PLIN2 (MsPLIN2), and studied developmental and nutritional changes in the expression of PLIN2. Nutritional changes induced fast alterations in PLIN2 mRNA and protein levels in fat body and midgut of the feeding larvae. The relationship observed between PLIN2 expression and TG synthesis in both larval fat body and midgut suggests that PLIN2 is needed when tissues are accumulating TG. However, when the fat body was storing TG at maximal capacity, MsPLIN2 levels declined. This unexpected finding suggests the occurrence of alternative mechanism/s to shield TG from the action of lipases in M. sexta LDs. In addition, it implies that the cellular level of lipid storage could be modulating MsPLIN2 expression and/or degradation. The study also confirmed that MsPLIN2 was most abundant in the adult fat body, which is characterized by a high rate TG hydrolysis and lipid mobilization. Whether MsPLIN2 is directly involved in lipolysis and/or the secretion of lipids in the fat body of adult of M. sexta is unknown at this time. Nonetheless, the coexistence of high PLIN2 level and lipolysis suggests a complex role for MsPLIN2. Altogether, we found that MsPLIN2 is needed when the synthesis of glycerides, DG and TG, is active even if the insect is accumulating or consuming TG.

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Section: Resultssupporting

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Clues on the function of Manduca sexta perilipin 2 inferred from developmental and nutrition-dependent changes in its expression

Chen

Firdaus

Howard

et al. 2017

Insect Biochemistry and Molecular Biology

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“…For Klar, targeting to lipid droplets is mediated by its C-terminal LD-domain, possibly via an amphipathic helix [56, 105]. LSD-2 binding to droplets is dominated by hydrophobic interactions [15], and a model for droplet binding has been proposed by modeling and analysis of GST fusions [148]. …”

Section: The Droplet Proteomementioning

confidence: 99%

As the fat flies: The dynamic lipid droplets of Drosophila embryos

Welte

2015

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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Research into lipid droplets is rapidly expanding, and new cellular and organismal roles for these lipid storage organelles are continually being discovered. The early Drosophila embryo is particularly well suited for addressing certain questions in lipid-droplet biology and combines technical advantages with unique biological phenomena. This review summarizes key features of this experimental system and the techniques available to study it, in order to make it accessible to researchers outside this field. It then describes the two topics most heavily studied in this system, lipid-droplet motility and protein sequestration on droplets, discusses what is known about the molecular players involved, points to open questions, and compares the results from Drosophila embryo studies to what it is known about lipid droplets in other systems.

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“…For example, packaging fat in intracellular lipid droplets and the mechanisms guiding mobilization of stored fat are conserved between mammals and insects (155)(156)(157)(158)(159). Lipid droplets, that are progressively recognized to represent ubiquitous dynamic organelles regulating intracellular TAG metabolism, are surrounded by a phospholipid monolayer coated with specific proteins, belonging to the evolutionary ancient PAT (perilipin/ADRP/TIP47) family of proteins, that participate in the regulation of TAG storage and lipolysis (156,157,(160)(161)(162)(163)(164)(165). For the lipid droplet-associated proteins in mammals PERILIPIN was recently adopted as a unifying nomenclature (165).…”

Section: Expert Opinionmentioning

confidence: 99%

Lipoprotein assembly and function in an evolutionary perspective

Horst

Rodenburg

2010

BioMolecular Concepts

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Circulatory fat transport in animals relies on members of the large lipid transfer protein (LLTP) superfamily, including mammalian apolipoprotein B (apoB) and insect apolipophorin II/I (apoLp-II/I). ApoB and apoLp-II/I, constituting the structural (non-exchangeable) basis for the assembly of various lipoproteins, acquire lipids through microsomal triglyceride-transfer protein, another LLTP family member, and bind them by means of amphipathic α-helical and β-sheet structural motifs. Comparative research reveals that LLTPs evolved from the earliest animals and highlights the structural adaptations in these lipid-binding proteins. Thus, in contrast to apoB, apoLp-II/I is cleaved post-translationally by a furin, resulting in the appearance of two non-exchangeable apolipoproteins in the single circulatory lipoprotein in insects, high-density lipophorin (HDLp). The remarkable structural similarities between mammalian and insect lipoproteins notwithstanding important functional differences relate to the mechanism of lipid delivery. Whereas in mammals, partial delipidation of apoB-containing lipoproteins eventually results in endocytic uptake of their remnants, mediated by members of the low-density lipoprotein receptor (LDLR) family, and degradation in lysosomes, insect HDLp functions as a reusable lipid shuttle capable of alternate unloading and reloading of lipid. Also, during muscular efforts (flight activity), an HDLp-based lipoprotein shuttle provides for the transport of lipid for energy generation. Although a lipophorin receptor - a homolog of LDLR - was identified that mediates endocytic uptake of HDLp during specific developmental periods, the endocytosed lipoprotein appears to be recycled in a transferrin-like manner. These data highlight that the functional adaptations in the lipoprotein lipid carriers in mammals and insects also emerge with regard to the functioning of their cognate receptors.

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Purification and Characterization of Recombinant Lipid Storage Protein-2 from Drosophila melanogaster

Cited by 21 publications

References 25 publications

Clues on the function of Manduca sexta perilipin 2 inferred from developmental and nutrition-dependent changes in its expression

Clues on the function of Manduca sexta perilipin 2 inferred from developmental and nutrition-dependent changes in its expression

As the fat flies: The dynamic lipid droplets of Drosophila embryos

Lipoprotein assembly and function in an evolutionary perspective

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