PPARδ activation induces hepatic long-chain acyl-CoA synthetase 4 expression in vivo and in vitro

Seed oil from flax (Linum usitatissimum) is enriched in α-linolenic acid (ALA; 18:3Δ9cis,12cis,15cis), but the biochemical processes underlying the enrichment of flax seed oil with this polyunsaturated fatty acid are not fully elucidated. Here, a potential process involving the catalytic actions of long-chain acyl-CoA synthetase (LACS) and diacylglycerol acyltransferase (DGAT) is proposed for ALA enrichment in triacylglycerol (TAG). LACS catalyzes the ATP-dependent activation of free fatty acid to form acyl-CoA, which in turn may serve as an acyl-donor in the DGAT-catalyzed reaction leading to TAG. To test this hypothesis, flax LACS and DGAT cDNAs were functionally expressed in Saccharomyces cerevisiae strains to probe their possible involvement in the enrichment of TAG with ALA. Among the identified flax LACSs, LuLACS8A exhibited significantly enhanced specificity for ALA over oleic acid (18:1Δ9cis) or linoleic acid (18:2Δ9cis,12cis). Enhanced α-linolenoyl-CoA specificity was also observed in the enzymatic assay of flax DGAT2 (LuDGAT2-3), which displayed ∼20 times increased preference toward α-linolenoyl-CoA over oleoyl-CoA. Moreover, when LuLACS8A and LuDGAT2-3 were co-expressed in yeast, both in vitro and in vivo experiments indicated that the ALA-containing TAG enrichment process was operative between LuLACS8A- and LuDGAT2-3-catalyzed reactions. Overall, the results support the hypothesis that the cooperation between the reactions catalyzed by LACS8 and DGAT2 may represent a route to enrich ALA production in the flax seed oil.

Download Full-text

IN VIVO AND IN VITRO ANALYSIS OF THE HUMAN LONG CHAIN ACYL COA DEHYDROGENASE(LCAD) GENE PROMOTER REGION. † 889

Pediatric Research ◽

10.1203/00006450-199604001-00911 ◽

1996 ◽

Vol 39 ◽

pp. 151-151

Author(s):

Zhifang Zhang ◽

Yeqing Zhou ◽

Arnold W Strauss

Keyword(s):

Promoter Region ◽

Gene Promoter ◽

Chain Acyl ◽

Vitro Analysis ◽

Gene Promoter Region ◽

Long Chain ◽

In Vitro Analysis

Download Full-text

In vitro characterization and in vivo expression of human very-long chain acyl-CoA dehydrogenase

Molecular Genetics and Metabolism ◽

10.1016/j.ymgme.2006.02.010 ◽

2006 ◽

Vol 88 (4) ◽

pp. 351-358 ◽

Cited By ~ 6

Author(s):

J. Lawrence Merritt ◽

Dietrich Matern ◽

Jerry Vockley ◽

Jan Daniels ◽

Tien V. Nguyen ◽

...

Keyword(s):

In Vitro Characterization ◽

In Vivo Expression ◽

Chain Acyl ◽

Long Chain

Download Full-text

Demonstration and characterization of phosphate transport in mammalian peroxisomes

Biochemical Journal ◽

10.1042/bj20041846 ◽

2005 ◽

Vol 389 (3) ◽

pp. 717-722 ◽

Cited By ~ 14

Author(s):

Wouter F. Visser ◽

Carlo W. van Roermund ◽

Lodewijk Ijlst ◽

Klaas J. Hellingwerf ◽

Ronald J. A. Wanders ◽

...

Keyword(s):

Small Molecules ◽

Adenine Nucleotide ◽

Phosphate Transport ◽

Significant Extent ◽

Peroxisomal Membrane ◽

Chain Acyl ◽

Long Chain

It is now well established that the peroxisomal membrane is not freely permeable to small molecules in vivo, which implies the existence of metabolite transporters in the peroxisomal membrane. A few putative peroxisomal metabolite transporters have indeed been identified, but the function of these proteins has remained largely unresolved so far. The only peroxisomal transporter characterized to a significant extent is the adenine nucleotide transporter, which is presumably required to sustain the activity of the intraperoxisomal very-long-chain-acyl-CoA synthetase. In addition to AMP, this acyl-CoA synthetase also produces pyrophosphate, which must be exported from the peroxisome. In the present study, we demonstrate that the peroxisomal membrane contains a transporter activity that facilitates the passage of phosphate and possibly pyrophosphate across the peroxisomal membrane. By reconstitution of peroxisomal membrane proteins in proteoliposomes, some kinetic parameters of the transporter could be established in vitro. The transporter can be distinguished from the mitochondrial phosphate transporter by its different sensitivity to inhibitors.

Download Full-text

Glucose Oligosaccharide and Long-Chain Glucomannan Feed Additives Induce Enhanced Activation of Intraepithelial NK Cells and Relative Abundance of Commensal Lactic Acid Bacteria in Broiler Chickens

Veterinary Sciences ◽

10.3390/vetsci8060110 ◽

2021 ◽

Vol 8 (6) ◽

pp. 110

Author(s):

Nathalie Meijerink ◽

Jean E. de Oliveira ◽

Daphne A. van Haarlem ◽

Guilherme Hosotani ◽

David M. Lamot ◽

...

Keyword(s):

Immune System ◽

Nk Cells ◽

Relative Abundance ◽

Nk Cell ◽

Feed Additives ◽

Microbiota Composition ◽

In Ovo ◽

Long Chain

Restrictions on the use of antibiotics in the poultry industry stimulate the development of alternative nutritional solutions to maintain or improve poultry health. This requires more insight in the modulatory effects of feed additives on the immune system and microbiota composition. Compounds known to influence the innate immune system and microbiota composition were selected and screened in vitro, in ovo, and in vivo. Among all compounds, 57 enhanced NK cell activation, 56 increased phagocytosis, and 22 increased NO production of the macrophage cell line HD11 in vitro. Based on these results, availability and regulatory status, six compounds were selected for further analysis. None of these compounds showed negative effects on growth, hatchability, and feed conversion in in ovo and in vivo studies. Based on the most interesting numerical results and highest future potential feasibility, two compounds were analyzed further. Administration of glucose oligosaccharide and long-chain glucomannan in vivo both enhanced activation of intraepithelial NK cells and led to increased relative abundance of lactic acid bacteria (LAB) amongst ileum and ceca microbiota after seven days of supplementation. Positive correlations between NK cell subsets and activation, and relative abundance of LAB suggest the involvement of microbiota in the modulation of the function of intraepithelial NK cells. This study identifies glucose oligosaccharide and long-chain glucomannan supplementation as effective nutritional strategies to modulate the intestinal microbiota composition and strengthen the intraepithelial innate immune system.

Download Full-text

Effect of long-chain fatty acyl-CoA on mitochondrial and cytosolic ATP/ADP ratios in the intact liver cell

Biochemical Journal ◽

10.1042/bj2200371 ◽

1984 ◽

Vol 220 (2) ◽

pp. 371-376 ◽

Cited By ~ 40

Author(s):

S Soboll ◽

H J Seitz ◽

H Sies ◽

B Ziegler ◽

R Scholz

Keyword(s):

Liver Cell ◽

Adenine Nucleotide ◽

Intact Cell ◽

Inhibitory Effect ◽

Fatty Acyl ◽

Fed State ◽

Physiological Relevance ◽

Chain Acyl ◽

Long Chain

The effect of long-chain acyl-CoA on subcellular adenine nucleotide systems was studied in the intact liver cell. Long-chain acyl-CoA content was varied by varying the nutritional state (fed and starved states) or by addition of oleate. Starvation led to an increase in the mitochondrial and a decrease in the cytosolic ATP/ADP ratio in liver both in vivo and in the isolated perfused organ as compared with the fed state. The changes were reversed on re-feeding glucose in liver in vivo or on infusion of substrates (glucose, glycerol) in the perfused liver, respectively. Similar changes in mitochondrial and cytosolic ATP/ADP ratios occurred on addition of oleate, but, importantly, not with a short-chain fatty acid such as octanoate. It is concluded that long-chain acyl-CoA exerts an inhibitory effect on mitochondrial adenine nucleotide translocation in the intact cell, as was previously postulated in the literature from data obtained with isolated mitochondria. The physiological relevance with respect to pyruvate metabolism, i.e. regulation of pyruvate carboxylase and pyruvate dehydrogenase by the mitochondrial ATP/ADP ratio, is discussed.

Download Full-text

Factor XII Activation Promotes Platelet Consumption in the Presence of Bacterial-Type Long-Chain Polyphosphate In Vitro and In Vivo

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.118.311193 ◽

2018 ◽

Vol 38 (8) ◽

pp. 1748-1760 ◽

Cited By ~ 15

Author(s):

Jevgenia Zilberman-Rudenko ◽

Stéphanie E. Reitsma ◽

Cristina Puy ◽

Rachel A. Rigg ◽

Stephanie A. Smith ◽

...

Keyword(s):

Factor Xii ◽

Bacterial Type ◽

Long Chain

Download Full-text

Phosphatidylcholine Synthesis Influences the Diacylglycerol Homeostasis Required for Sec14p-dependent Golgi Function and Cell Growth

Molecular Biology of the Cell ◽

10.1091/mbc.12.3.511 ◽

2001 ◽

Vol 12 (3) ◽

pp. 511-520 ◽

Cited By ~ 48

Author(s):

Annette L. Henneberry ◽

Thomas A. Lagace ◽

Neale D. Ridgway ◽

Christopher R. McMaster

Keyword(s):

Cell Death ◽

Cell Growth ◽

Phosphatidic Acid ◽

Metabolic Pathways ◽

Eukaryotic Cells ◽

Kennedy Pathway ◽

Phosphatidylcholine Synthesis ◽

Long Chain

Phosphatidylcholine and phosphatidylethanolamine are the most abundant phospholipids in eukaryotic cells and thus have major roles in the formation and maintenance of vesicular membranes. In yeast, diacylglycerol accepts a phosphocholine moiety through aCPT1-derived cholinephosphotransferase activity to directly synthesize phosphatidylcholine. EPT1-derived activity can transfer either phosphocholine or phosphoethanolamine to diacylglcyerol in vitro, but is currently believed to primarily synthesize phosphatidylethanolamine in vivo. In this study we report that CPT1- and EPT1-derived cholinephosphotransferase activities can significantly overlap in vivo such that EPT1 can contribute to 60% of net phosphatidylcholine synthesis via the Kennedy pathway. Alterations in the level of diacylglycerol consumption through alterations in phosphatidylcholine synthesis directly correlated with the level of SEC14-dependent invertase secretion and affected cell viability. Administration of synthetic di8:0 diacylglycerol resulted in a partial rescue of cells fromSEC14-mediated cell death. The addition of di8:0 diacylglycerol increased di8:0 diacylglycerol levels 20–40-fold over endogenous long-chain diacylglycerol levels. Di8:0 diacylglcyerol did not alter endogenous phospholipid metabolic pathways, nor was it converted to di8:0 phosphatidic acid.

Download Full-text