scholarly journals Isolation and transcriptomic analysis of Anopheles gambiae oenocytes enables the delineation of hydrocarbon biosynthesis

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Linda Grigoraki ◽  
Xavier Grau-Bové ◽  
Henrietta Carrington Yates ◽  
Gareth J Lycett ◽  
Hilary Ranson
Keyword(s):  
Anopheles Gambiae ◽  
Biosynthetic Pathway ◽  
Saturated Fatty Acids ◽  
Fatty Acyl ◽  
Arid Environments ◽  
Biological Processes ◽  
Ecological Habitat ◽  
First Time ◽  
Hydrocarbon Biosynthesis ◽  
Long Chain

The surface of insects is coated in cuticular hydrocarbons (CHCs); variations in the composition of this layer affect a range of traits including adaptation to arid environments and defence against pathogens and toxins. In the African malaria vector, Anopheles gambiae quantitative and qualitative variance in CHC composition have been associated with speciation, ecological habitat and insecticide resistance. Understanding how these modifications arise will inform us of how mosquitoes are responding to climate change and vector control interventions. CHCs are synthesised in sub-epidermal cells called oenocytes that are very difficult to isolate from surrounding tissues. Here we utilise a transgenic line with fluorescent oenocytes to purify these cells for the first time. Comparative transcriptomics revealed the enrichment of biological processes related to long chain fatty acyl-CoA biosynthesis and elongation of mono-, poly-unsaturated and saturated fatty acids and enabled us to delineate, and partially validate, the hydrocarbon biosynthetic pathway in An. gambiae.

scholarly journals Cuticular hydrocarbon biosynthesis in malaria vectors: insights from the adult oenocyte transcriptome

2020 ◽  
Author(s):  
Linda Grigoraki ◽  
Xavier Grau-Bove ◽  
Henrietta Carrington-Yates ◽  
Gareth J Lycett ◽  
Hilary Ranson
Keyword(s):  
Biosynthetic Pathway ◽  
Saturated Fatty Acids ◽  
Cuticular Hydrocarbon ◽  
Fatty Acyl ◽  
Surrounding Tissue ◽  
Malaria Vectors ◽  
Arid Environments ◽  
Ecological Habitat ◽  
First Time ◽  
Hydrocarbon Biosynthesis

AbstractThe surface of insects is coated in cuticular hydrocarbons (CHCs); variations in the composition of this layer affect a range of traits including adaptation to arid environments and defence against pathogens and toxins. In the African malaria vector, Anopheles gambiae quantitative and qualitative variance in CHC composition have been associated with speciation, ecological habitat and insecticide resistance. Understanding how these modifications arise will inform us of how mosquitoes are responding to climate change and vector control interventions. CHCs are synthesised in sub-epidermal cells called oenocytes that are very difficult to isolate from surrounding tissue. Here we utilise a transgenic line with fluorescent oenocytes to purify these cells for the first time. Comparative transcriptomics revealed the enrichment of biological processes related to long chain fatty acyl-CoA biosynthesis and elongation of mono-, poly-unsaturated and saturated fatty acids and enabled us to delineate, and partially validate, the hydrocarbon biosynthetic pathway in An gambiae.

scholarly journals Active Multienzyme Assemblies for Long-Chain Olefinic Hydrocarbon Biosynthesis

2017 ◽  
Vol 199 (9) ◽  
Author(s):  
James K. Christenson ◽  
Matthew R. Jensen ◽  
Brandon R. Goblirsch ◽  
Fatuma Mohamed ◽  
Wei Zhang ◽  
...  
Keyword(s):  
Active Sites ◽  
Xanthomonas Campestris ◽  
Fatty Acyl ◽  
Biosynthetic Activity ◽  
Content Type ◽  
Large Structures ◽  
Negative Stain ◽  
Acyl Coenzyme A ◽  
Hydrocarbon Biosynthesis ◽  
Long Chain

ABSTRACT Bacteria from different phyla produce long-chain olefinic hydrocarbons derived from an OleA-catalyzed Claisen condensation of two fatty acyl coenzyme A (acyl-CoA) substrates, followed by reduction and oxygen elimination reactions catalyzed by the proteins OleB, OleC, and OleD. In this report, OleA, OleB, OleC, and OleD were individually purified as soluble proteins, and all were found to be essential for reconstituting hydrocarbon biosynthesis. Recombinant coexpression of tagged OleABCD proteins from Xanthomonas campestris in Escherichia coli and purification over His6 and FLAG columns resulted in OleA separating, while OleBCD purified together, irrespective of which of the four Ole proteins were tagged. Hydrocarbon biosynthetic activity of copurified OleBCD assemblies could be reconstituted by adding separately purified OleA. Immunoblots of nondenaturing gels using anti-OleC reacted with X. campestris crude protein lysate indicated the presence of a large protein assembly containing OleC in the native host. Negative-stain electron microscopy of recombinant OleBCD revealed distinct large structures with diameters primarily between 24 and 40 nm. Assembling OleB, OleC, and OleD into a complex may be important to maintain stereochemical integrity of intermediates, facilitate the movement of hydrophobic metabolites between enzyme active sites, and protect the cell against the highly reactive β-lactone intermediate produced by the OleC-catalyzed reaction. IMPORTANCE Bacteria biosynthesize hydrophobic molecules to maintain a membrane, store carbon, and for antibiotics that help them survive in their niche. The hydrophobic compounds are often synthesized by a multidomain protein or by large multienzyme assemblies. The present study reports on the discovery that long-chain olefinic hydrocarbons made by bacteria from different phyla are produced by multienzyme assemblies in X. campestris. The OleBCD multienzyme assemblies are thought to compartmentalize and sequester olefin biosynthesis from the rest of the cell. This system provides additional insights into how bacteria control specific biosynthetic pathways.

scholarly journals Widespread Head-to-Head Hydrocarbon Biosynthesis in Bacteria and Role of OleA

2010 ◽  
Vol 76 (12) ◽  
pp. 3850-3862 ◽  
Author(s):  
David J. Sukovich ◽  
Jennifer L. Seffernick ◽  
Jack E. Richman ◽  
Jeffrey A. Gralnick ◽  
Lawrence P. Wackett
Keyword(s):  
Xanthomonas Campestris ◽  
Gene Deletion ◽  
Shewanella Oneidensis ◽  
Fatty Acyl ◽  
Double Bonds ◽  
Acyl Chains ◽  
Hydrocarbon Biosynthesis ◽  
Structural Alignments ◽  
Long Chain

ABSTRACT Previous studies identified the oleABCD genes involved in head-to-head olefinic hydrocarbon biosynthesis. The present study more fully defined the OleABCD protein families within the thiolase, α/β-hydrolase, AMP-dependent ligase/synthase, and short-chain dehydrogenase superfamilies, respectively. Only 0.1 to 1% of each superfamily represents likely Ole proteins. Sequence analysis based on structural alignments and gene context was used to identify highly likely ole genes. Selected microorganisms from the phyla Verucomicrobia, Planctomyces, Chloroflexi, Proteobacteria, and Actinobacteria were tested experimentally and shown to produce long-chain olefinic hydrocarbons. However, different species from the same genera sometimes lack the ole genes and fail to produce olefinic hydrocarbons. Overall, only 1.9% of 3,558 genomes analyzed showed clear evidence for containing ole genes. The type of olefins produced by different bacteria differed greatly with respect to the number of carbon-carbon double bonds. The greatest number of organisms surveyed biosynthesized a single long-chain olefin, 3,6,9,12,15,19,22,25,28-hentriacontanonaene, that contains nine double bonds. Xanthomonas campestris produced the greatest number of distinct olefin products, 15 compounds ranging in length from C28 to C31 and containing one to three double bonds. The type of long-chain product formed was shown to be dependent on the oleA gene in experiments with Shewanella oneidensis MR-1 ole gene deletion mutants containing native or heterologous oleA genes expressed in trans. A strain deleted in oleABCD and containing oleA in trans produced only ketones. Based on these observations, it was proposed that OleA catalyzes a nondecarboxylative thiolytic condensation of fatty acyl chains to generate a β-ketoacyl intermediate that can decarboxylate spontaneously to generate ketones.

Changes of dolichol and dolichyl fatty acyl esters during the germination and development of soybeans

1992 ◽  
Vol 70 (6) ◽  
pp. 466-469 ◽  
Author(s):  
Masataka Ishinaga ◽  
Toshimi Yamauchi ◽  
Keiko Egusa ◽  
Keiko Kuroda
Keyword(s):  
Fatty Acids ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Saturated Fatty Acids ◽  
Behenic Acid ◽  
Fatty Acyl ◽  
Short Chain ◽  
Soybean Seedlings ◽  
Long Chain

The amounts of dolichol and dolichyl fatty acyl esters and their composition in various parts of soybean seedlings were determined during germination and development. The dolichol content of cotyledons decreased during germination. Dolichyl fatty acyl esters were identified in cotyledons and the amount was estimated by high performance liquid chromatography. The relative amounts of short-chain dolichols of 15, 16, and 17 isoprene units increased during development of the seedlings. The homologue distribution of free dolichol was different from that of dolichyl fatty acyl esters. The relative amounts of dolichols with 16, 17, and 18 isoprene units were greater in free dolichol than in dolichyl fatty acyl esters. The percentages of long-chain saturated fatty acids in dolichyl fatty acyl esters, specifically 21:0, 22:0, 23:0, 24:0, and 25:0, increased during development. These fatty acids represented more than 40% of the fatty acids in dolichyl fatty acyl esters in stems. These results suggest that dolichyl fatty acyl esters are not a storage form of dolichol. The large accumulation of dolichol and dolichyl fatty acyl esters in the leaves, where photosynthesis takes place, suggests some other function.Key words: dolichol, dolichyl fatty acy ester, soybean, behenic acid, lignoceric acid.

scholarly journals Long-chain saturated fatty acids in breast milk are associated with the pathogenesis of atopic dermatitis via induction of inflammatory ILC3s

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Weng Sheng Kong ◽  
Naohiro Tsuyama ◽  
Hiroko Inoue ◽  
Yun Guo ◽  
Sho Mokuda ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Atopic Dermatitis ◽  
Breast Milk ◽  
Saturated Fatty Acids ◽  
Lymphoid Cells ◽  
Interleukin 17 ◽  
Immune System Development ◽  
Molecular Patterns ◽  
Damage Associated Molecular Patterns ◽  
Long Chain

AbstractBreastfeeding influences the immune system development in infants and may even affect various immunological responses later in life. Breast milk provides a rich source of early nutrition for infant growth and development. However, the presence of certain compounds in breast milk, related to an unhealthy lifestyle or the diet of lactating mothers, may negatively impact infants. Based on a cohort study of atopic dermatitis (AD), we find the presence of damage-associated molecular patterns (DAMPs) activity in the mother’s milk. By non-targeted metabolomic analysis, we identify the long-chain saturated fatty acids (LCSFA) as a biomarker DAMPs (+) breast milk samples. Similarly, a mouse model in which breastfed offspring are fed milk high in LCSFA show AD onset later in life. We prove that LCSFA are a type of damage-associated molecular patterns, which initiate a series of inflammatory events in the gut involving type 3 innate lymphoid cells (ILC3s). A remarkable increase in inflammatory ILC3s is observed in the gut, and the migration of these ILC3s to the skin may be potential triggers of AD. Gene expression analysis of ILC3s isolated from the gut reveal upregulation of genes that increase ILC3s and chemokines/chemokine receptors, which may play a role in ILC migration to the skin. Even in the absence of adaptive immunity, Rag1 knockout mice fed a high-LCSFA milk diet develop eczema, accompanied by increased gut ILC3s. We also present that gut microbiota of AD-prone PA milk-fed mice is different from non-AD OA/ND milk-fed mice. Here, we propose that early exposure to LCSFAs in infants may affect the balance of intestinal innate immunity, inducing a highly inflammatory environment with the proliferation of ILC3s and production of interleukin-17 and interleukin-22, these factors may be potential triggers or worsening factors of AD.

A Ratiometric Fluorescent Biosensor Reveals Dynamic Regulation of Long‐Chain Fatty Acyl‐CoA Esters Metabolism

2021 ◽  
Author(s):  
Jing Wang ◽  
Weibo Wang ◽  
Qingpeng Wei ◽  
Jiayuan Zhang ◽  
Meiqi Zhang ◽  
...  
Keyword(s):  
Fatty Acyl ◽  
Dynamic Regulation ◽  
Fluorescent Biosensor ◽  
Long Chain

scholarly journals Utility of measuring very long-chain fatty-acyl carnitines in dried blood spots for newborn screening of X-linked Adrenoleukodystrophy

2021 ◽  
Vol 26 ◽  
pp. 100720
Author(s):  
Archana Natarajan ◽  
Rita Christopher ◽  
Shruti V. Palakuzhiyil ◽  
Sadanandavalli Retnaswami Chandra
Keyword(s):  
Newborn Screening ◽  
Dried Blood Spots ◽  
Fatty Acyl ◽  
Blood Spots ◽  
Long Chain

Long chain fatty acyl-CoA modulation of H2O2 release at mitochondrial complex I

2008 ◽  
Vol 40 (1) ◽  
pp. 9-18 ◽  
Author(s):  
Silvia Bortolami ◽  
Evelin Comelato ◽  
Franco Zoccarato ◽  
Adolfo Alexandre ◽  
Lucia Cavallini
Keyword(s):  
Complex I ◽  
Fatty Acyl ◽  
Mitochondrial Complex I ◽  
Mitochondrial Complex ◽  
Long Chain

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

1984 ◽  
Vol 220 (2) ◽  
pp. 371-376 ◽  
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.

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