Very Long-chain Acyl-CoA Synthetases

Steinberg, Steven J.; Morgenthaler, Janine; Heinzer, Ann K.; Smith, Kirby D.; Watkins, Paul A.

doi:10.1074/jbc.m006403200

Cited by 122 publications

(64 citation statements)

References 30 publications

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“…This distribution together with the fact that, among the fatty acids tested, only hexadecanoic acid and octadecanoic acid competitively inhibited the activation of 2-hydroxyoctadecanoic acid, point toward the long chain acylCoA synthetase as the activating enzyme. However, because it is well known that the substrate spectra of long chain and very long chain acyl-CoA synthetases overlap (39,40), no definitive conclusions can be drawn as to the specific enzyme involved. Moreover, it might well be that the chain length of the substrate determines the involved isoform.…”

Section: Discussionmentioning

confidence: 98%

Breakdown of 2-Hydroxylated Straight Chain Fatty Acids via Peroxisomal 2-Hydroxyphytanoyl-CoA Lyase

Foulon

Sniekers

Huysmans

et al. 2005

Journal of Biological Chemistry

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2-Hydroxyfatty acids, constituents of brain cerebrosides and sulfatides, were previously reported to be degraded by an ␣-oxidation system, generating fatty acids shortened by one carbon atom. In the current study we used labeled and unlabeled 2-hydroxyoctadecanoic acid to reinvestigate the degradation of this class of lipids. Both in intact and broken cell systems formate was identified as a main reaction product. Furthermore, the generation of an n ؊ 1 aldehyde was demonstrated. In permeabilized rat hepatocytes and liver homogenates, studies on cofactor requirements revealed a dependence on ATP, CoA, Mg 2؉ , thiamine pyrophosphate, and NAD ؉ . Together with subcellular fractionation data and studies on recombinant enzymes, this led to the following picture. In a first step, the 2-hydroxyfatty acid is activated to an acyl-CoA; subsequently, the 2-hydroxy fatty acyl-CoA is cleaved by 2-hydroxyphytanoyl-CoA lyase, to formyl-CoA and an n ؊ 1 aldehyde. The severe inhibition of formate generation by oxythiamin treatment of intact fibroblasts indicates that cleavage through the thiamine pyrophosphate-dependent 2-hydroxyphytanoyl-CoA lyase is the main pathway for the degradation of 2-hydroxyfatty acids. The latter protein was initially characterized as an essential enzyme in the peroxisomal ␣-oxidation of 3-methyl-branched fatty acids such as phytanic acid. Our findings point to a new role for peroxisomes in mammals, i.e. the breakdown of 2-hydroxyfatty acids, at least the long chain 2-hydroxyfatty acids. Most likely, the more abundant very long chain 2-hydroxyfatty acids are degraded in a similar manner.

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Section: Discussionmentioning

confidence: 98%

Breakdown of 2-Hydroxylated Straight Chain Fatty Acids via Peroxisomal 2-Hydroxyphytanoyl-CoA Lyase

Foulon

Sniekers

Huysmans

et al. 2005

Journal of Biological Chemistry

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“…Oxidation of [1][2][3][4][5][6][7][8][9][10][11][12][13][14] C] fatty acids to water-soluble products was also measured as previously described (27). Briefly, fatty acids were solubilized with ␣-cyclodextrin as described for ACS assays and incubated in 20 mM Tris, pH 8.0, for 1 h at 37°C with freshly harvested cell suspensions and required cofactors (8.5 mM ATP, 8.5 mM MgCl 2 , 1 mM NAD, 0.17 mM FAD, 2.5 mM carnitine, 0.16 mM CoA, 1 mM malate).…”

Section: Methodsmentioning

confidence: 99%

“…A 108-bp fragment was amplified by PCR using FATP4 full-length cDNA 5 as template with forward primer 5Ј-ATGCCGAATTCCGCTATTGTGAAAGAC-CCG-3Ј, which incorporates an EcoRI site (underlined), and reverse primer 5Ј-TATTCTCGAGTCACAGCTTCTCCTCG-CCTGC-3Ј, which incorporates an XhoI site (underlined) just after the TGA stop codon (reverse complement shown in bold). This fragment was cloned in frame into the EcoRI and XhoI sites of the pGEX-His-T3 vector (Amersham Biosciences).…”

Section: Methodsmentioning

confidence: 99%

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Fatty Acid Transport Protein 4 Is the Principal Very Long Chain Fatty Acyl-CoA Synthetase in Skin Fibroblasts

Jia

Moulson

Pei

et al. 2007

Journal of Biological Chemistry

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102

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Fatty acid transport protein 4 (FATP4) is a fatty acyl-CoA synthetase that preferentially activates very long chain fatty acid substrates, such as C24:0, to their CoA derivatives. To gain better insight into the physiological functions of FATP4, we established dermal fibroblast cell lines from FATP4-deficient wrinkle-free mice and wild type (w.t.) mice. FATP4 ؊/؊ fibroblasts had no detectable FATP4 protein by Western blot. Compared with w.t. fibroblasts, cells lacking FATP4 had an 83% decrease in C24:0 activation. Peroxisomal degradation of C24:0 was reduced by 58%, and rates of C24:0 incorporation into major phospholipid species (54 -64% decrease), triacylglycerol (64% decrease), and cholesterol esters (58% decrease) were significantly diminished. Because these lipid metabolic processes take place in different subcellular organelles, we used immunofluorescence and Western blotting of subcellular fractions to investigate the distribution of FATP4 protein and measured enzyme activity in fractions from w.t. and FATP4 ؊/؊ fibroblasts. FATP4 protein and acyl-CoA synthetase activity localized to multiple organelles, including mitochondria, peroxisomes, endoplasmic reticulum, and the mitochondria-associated membrane fraction. We conclude that in murine skin fibroblasts, FATP4 is the major enzyme producing very long chain fatty acid-CoA for lipid metabolic pathways. Although FATP4 deficiency primarily affected very long chain fatty acid metabolism, mutant fibroblasts also showed reduced uptake of a fluorescent long chain fatty acid and reduced levels of long chain polyunsaturated fatty acids. FATP4-deficient cells also contained abnormal neutral lipid droplets. These additional defects indicate that metabolic abnormalities in these cells are not limited to very long chain fatty acids.Fatty acids containing 22 or more carbon atoms are referred to as very long chain fatty acids (VLCFA).2 VLCFA are normal constituents of membrane lipids but are particularly abundant in brain, testis, and skin (1-3). Fatty acids, including VLCFA, cannot participate in most metabolic processes unless they are first activated to their CoA derivatives (4). Once activated, however, fatty acids can be degraded to release stored energy or incorporated into glycerolipids, phospholipids, sphingolipids, glycolipids, cholesterol esters, and other complex lipids. Certain acyl-CoAs also serve as substrates for protein acylation and/or function as regulatory molecules. Thus, the fatty acid activation reaction is central to normal cellular lipid metabolism.The acyl-CoA synthetase (ACS) enzyme family catalyzes this essential reaction (4). Because fatty acid chain lengths vary from 2 to more than 30 carbon atoms, different ACSs capable of activating short, medium, long, and very long chain fatty acids have evolved. Based on analysis of amino acid sequence homology, we estimate that mammalian genomes encode at least 25 different ACSs.3 Further analysis of highly conserved domains within these sequences allowed us to group the enzymes into families (5). We ...

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“…A second conserved motif (fatty acyl-CoA synthetase [FACS] signature motif) has been identified in ACSs activating long-chain FAs (15). Variations of the FACS motif have been described for ACSs activating other length classes of FA: medium chain (16), very long chain (17), and bubblegum (18). Structural determination of a longchain ACS from the bacterium Thermus thermophilus provided insights into four conserved regions in fatty ACSs: adenine binding, linker, and gate motifs, as well as the phosphate binding site which corresponds to motif I (19).…”

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confidence: 99%

Physiological Role of Acyl Coenzyme A Synthetase Homologs in Lipid Metabolism in Neurospora crassa

et al. 2013

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Acyl coenzyme A (CoA) synthetase (ACS) enzymes catalyze the activation of free fatty acids (FAs) to CoA esters by a two-step thioesterification reaction. Activated FAs participate in a variety of anabolic and catabolic lipid metabolic pathways, including de novo complex lipid biosynthesis, FA ␤-oxidation, and lipid membrane remodeling. Analysis of the genome sequence of the filamentous fungus Neurospora crassa identified seven putative fatty ACSs (ACS-1 through ACS-7). ACS-3 was found to be the major activator for exogenous FAs for anabolic lipid metabolic pathways, and consistent with this finding, ACS-3 localized to the endoplasmic reticulum, plasma membrane, and septa. Double-mutant analyses confirmed partial functional redundancy of ACS-2 and ACS-3. ACS-5 was determined to function in siderophore biosynthesis, indicating alternative functions for ACS enzymes in addition to fatty acid metabolism. The N. crassa ACSs involved in activation of FAs for catabolism were not specifically defined, presumably due to functional redundancy of several of ACSs for catabolism of exogenous FAs. Lipids are essential components of all living cells. They are major components of biological membranes (e.g., phospholipids) and serve as energy reserves (e.g., triacylglycerols) as well as signaling molecules (e.g., sphingolipids and lysophosphatidic acid). In fungi, fatty acids (FAs) are synthesized de novo via the FA synthase complex (FAS type I) present in the cytosol (1) or in the mitochondria via a suite of enzymes, including an acyl carrier protein. These mitochondrial FAS type II enzymes are not present in a complex (2). Many fungi can also utilize exogenous FAs, both for incorporation into lipids and as a carbon source. In addition to incorporation into complex lipids in the endoplasmic reticulum (ER) (3, 4) and mitochondria (5), FAs can be remodeled by desaturation and elongation (6-8) and degraded for energy production in the peroxisome (glyoxysome) (9, 10) and in the mitochondria (9, 11).One of the most important catalytic activities in lipid metabolism is activation of FAs by acyl coenzyme A (CoA) synthetase (ACS) (EC 6.2.1.3). Activated FAs participate in a number of cellular metabolic pathways, including synthesis of phospholipids, triacylglycerols, and cholesterol esters, FA elongation, FA desaturation, and ␤-oxidation. ACSs catalyze the two-step ATP-dependent reaction of FA activation, whereby first the FA substrate is adenylated to form an acyl-AMP intermediate and subsequently AMP is exchanged with CoA, forming a thioester bond to yield the activated acyl-CoA (12).The number of carbons present in the FA substrate of ACSs ranges from 2 to more than 26. The characteristic length of the FA substrate defines subfamilies of ACSs as short-chain (SC; 2 to 4 carbons), medium-chain (MC; 4 to 12 carbons), long-chain (LC; 12 to 20 carbons), very-long-chain (VLC; 18 to 26 carbons), and "bubblegum" (14 to 24 carbons) FA activators (13). All ACSs contain a highly conserved ATP/AMP binding motif (14), which is conserved among all...

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Very Long-chain Acyl-CoA Synthetases

Cited by 122 publications

References 30 publications

Breakdown of 2-Hydroxylated Straight Chain Fatty Acids via Peroxisomal 2-Hydroxyphytanoyl-CoA Lyase

Breakdown of 2-Hydroxylated Straight Chain Fatty Acids via Peroxisomal 2-Hydroxyphytanoyl-CoA Lyase

Fatty Acid Transport Protein 4 Is the Principal Very Long Chain Fatty Acyl-CoA Synthetase in Skin Fibroblasts

Physiological Role of Acyl Coenzyme A Synthetase Homologs in Lipid Metabolism in Neurospora crassa

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