Die Synthese der Fettsäuren mit gereinigten Enzymen des Fettsäurecyclus

Seubert, W.; Greull, Gerhard; Lynen, Feodor

doi:10.1002/ange.19570691103

Cited by 180 publications

(22 citation statements)

References 8 publications

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“…The holo-ACP-A or holo-PCP were separated from phosphopantetheinyltransferase by nickel ion affinity chromatography. Confirmation that at least 90% of the apoforms had been converted to the corresponding holoforms was obtained by incubation of a portion of the product with [1][2][3][4][5][6][7][8][9][10][11][12][13][14] C]acetyl-CoA and human phosphopantetheinyltransferase and assaying the incorporation of radiolabel into the ACP (only residual apoform is radiolabeled by [1][2][3][4][5][6][7][8][9][10][11][12][13][14] C]acetyl-phosphopantetheine transfer (24)). …”

Section: Subcellular Fractionation Of Sf9 Cells Expressing the Full-lmentioning

confidence: 99%

“…Indeed one laboratory reported that short and medium chain-length fatty acids could be produced using the partially purified mitochondrial ␤-oxidation enzymes, ␤-ketoacyl thiolase, ␤-hydroxyacyl-CoA dehydrogenase, enoyl-CoA hydratase, and enoyl-CoA reductase (4), and another found that inhibitory antibodies to pig heart 3-ketoacyl-CoA thiolase inhibited, in a parallel fashion, the fatty acid elongation system of pig heart mitochondria (5). The picture was complicated by reports that liver and heart mitochondria have two distinct fatty acid-synthesizing systems, one acetyl-CoA-dependent, possibly involving some of the ␤-oxidation enzymes, the other malonyl-CoA-dependent and apparently resembling the cytosolic FAS 1 system. Most studies were in agreement that the acetyl-CoA-dependent system was an elongation pathway in which one or more C 2 units, derived from acetyl-CoA, were added to preexisting saturated and unsaturated fatty acids of a broad range of chain length (C 6 -C 20 ).…”

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confidence: 99%

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Cloning, Expression, Characterization, and Interaction of Two Components of a Human Mitochondrial Fatty Acid Synthase

Zhang¹,

Joshi²,

Smith³

2003

Journal of Biological Chemistry

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The possibility that human cells contain, in addition to the cytosolic type I fatty acid synthase complex, a mitochondrial type II malonyl-CoA-dependent system for the biosynthesis of fatty acids has been examined by cloning, expressing, and characterizing two putative components. Candidate coding sequences for a malonylCoA:acyl carrier protein transacylase (malonyltransferase) and its acyl carrier protein substrate, identified by BLAST searches of the human sequence data base, were located on nuclear chromosomes 22 and 16, respectively. The encoded proteins localized exclusively in mitochondria only when the putative N-terminal mitochondrial targeting sequences were present as revealed by confocal microscopy of HeLa cells infected with appropriate green fluorescent protein fusion constructs. The mature, processed forms of the mitochondrial proteins were expressed in Sf9 cells and purified, the acyl carrier protein was converted to the holoform in vitro using purified human phosphopantetheinyltransferase, and the functional interaction of the two proteins was studied. Compared with the dual specificity malonyl/ acetyltransferase component of the cytosolic type I fatty acid synthase, the type II mitochondrial counterpart exhibits a relatively narrow substrate specificity for both the acyl donor and acyl carrier protein acceptor. Thus, it forms a covalent acyl-enzyme complex only when incubated with malonyl-CoA and transfers exclusively malonyl moieties to the mitochondrial holoacyl carrier protein. The type II acyl carrier protein from Bacillus subtilis, but not the acyl carrier protein derived from the human cytosolic type I fatty acid synthase, can also function as an acceptor for the mitochondrial transferase. These data provide compelling evidence that human mitochondria contain a malonylCoA/acyl carrier protein-dependent fatty acid synthase system, distinct from the type I cytosolic fatty acid synthase, that resembles the type II system present in prokaryotes and plastids. The final products of this system, yet to be identified, may play an important role in mitochondrial function.The existence of a mitochondrial system for the biosynthesis of fatty acids in animals was first reported 40 years ago and postulated to function by a reversal of the process of fatty acid ␤-oxidation (1-3). Indeed one laboratory reported that short and medium chain-length fatty acids could be produced using the partially purified mitochondrial ␤-oxidation enzymes, ␤-ketoacyl thiolase, ␤-hydroxyacyl-CoA dehydrogenase, enoyl-CoA hydratase, and enoyl-CoA reductase (4), and another found that inhibitory antibodies to pig heart 3-ketoacyl-CoA thiolase inhibited, in a parallel fashion, the fatty acid elongation system of pig heart mitochondria (5). The picture was complicated by reports that liver and heart mitochondria have two distinct fatty acid-synthesizing systems, one acetyl-CoA-dependent, possibly involving some of the ␤-oxidation enzymes, the other malonyl-CoA-dependent and apparently resembling the cytosolic FAS 1 system. Most ...

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Section: Subcellular Fractionation Of Sf9 Cells Expressing the Full-lmentioning

confidence: 99%

mentioning

confidence: 99%

Cloning, Expression, Characterization, and Interaction of Two Components of a Human Mitochondrial Fatty Acid Synthase

Zhang¹,

Joshi²,

Smith³

2003

Journal of Biological Chemistry

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“…But after treatment of the homogenate with deoxycholic acid [23], structural enzymes may be dispersed and assayed by the more convenient and rapid optical assay according to Warburg [24]. This method was also applied to assays of the transport ATPase.…”

Section: Assaymentioning

confidence: 99%

On the Mechanism of Na⁺‐and K⁺‐Stimulated Hydrolysis of Adenosine Triphosphate

Schoner

Ilberg

Kramer

et al. 1967

European Journal of Biochemistry

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265

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“…Cholesterol (Bucher & McGarrahan, 1956) and fatty acids (Langdon, 1955;Seubert, Greull & Lynen, 1957) have been shown to be synthesized in vitro by homogenates of mammalian liver from which the heavier subcellular particles have been removed. We have therefore studied cholesterol and fatty acid synthesis in such preparations, obtained from the livers of thyroxine-treated and normal rats.…”

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confidence: 99%

Effects of thyroxine on the synthesis of cholesterol and fatty acids by cell‐free fractions of rat liver

Fletcher¹,

Myant²

1960

The Journal of Physiology

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We have shown elsewhere (Fletcher & Myant, 1958) that the synthesis of cholesterol from acetate by rat liver slices is increased if the rat is given injections of thyroxine before the liver is removed. If mevalonic acid, instead of acetate, is used as the precursor of cholesterol, this effect is absent or only just detectable. When the daily dose of thyroxine is 20 ,ug or more, synthesis of fatty acids by the liver slices is depressed. It was suggested that treatment with thyroxine stimulates cholesterol synthesis by acting upon a rate-limiting step between acetate and mevalonic acid, and that the depression of fatty acid synthesis might be due to a deficiency of ATP.In this paper we describe experiments designed to test whether these effects of thyroxine can be localized in one or other of the fractions of the liver cell. Cholesterol (Bucher & McGarrahan, 1956) and fatty acids (Langdon, 1955;Seubert, Greull & Lynen, 1957) have been shown to be synthesized in vitro by homogenates of mammalian liver from which the heavier subcellular particles have been removed. We have therefore studied cholesterol and fatty acid synthesis in such preparations, obtained from the livers of thyroxine-treated and normal rats. METHODSAnimals. The rats were albino males of an inbred strain, weighing 120-200 g. The treated animals were given sodium-L-thyroxine (20-50 jg in 0-2 ml. water) daily for 9 days by intraperitoneal injection under light ether anaesthesia. The solutions of thyroxine were made by dissolving the required amount in a small quantity of water with the addition of one drop of 20 % NaOH, and then diluting to 100 ml. with water. Control animals were similarly injected with 0-2 ml. of an alkaline solution containing no thyroxine. During treatment with 20 or 30 ,g of thyroxine per day, the weight gain and food consumption of the treated rats did not differ significantly from that of the controls. When the daily dose of thyroxine was 50 Zg, the treated animals gained less weight than the controls.Preparation of tissue. At the end of the treatment, the animals were killed by breaking their necks. The livers were removed as rapidly as possible and placed on ice. Homogenates were then prepared from the pooled livers of each group of treated and control rats, 10PHYSIO. CLIV

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Die Synthese der Fettsäuren mit gereinigten Enzymen des Fettsäurecyclus

Cited by 180 publications

References 8 publications

Cloning, Expression, Characterization, and Interaction of Two Components of a Human Mitochondrial Fatty Acid Synthase

Cloning, Expression, Characterization, and Interaction of Two Components of a Human Mitochondrial Fatty Acid Synthase

On the Mechanism of Na⁺‐and K⁺‐Stimulated Hydrolysis of Adenosine Triphosphate

Effects of thyroxine on the synthesis of cholesterol and fatty acids by cell‐free fractions of rat liver

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Die Synthese der Fettsäuren mit gereinigten Enzymen des Fettsäurecyclus

Cited by 180 publications

References 8 publications

Cloning, Expression, Characterization, and Interaction of Two Components of a Human Mitochondrial Fatty Acid Synthase

Cloning, Expression, Characterization, and Interaction of Two Components of a Human Mitochondrial Fatty Acid Synthase

On the Mechanism of Na+‐and K+‐Stimulated Hydrolysis of Adenosine Triphosphate

Effects of thyroxine on the synthesis of cholesterol and fatty acids by cell‐free fractions of rat liver

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On the Mechanism of Na⁺‐and K⁺‐Stimulated Hydrolysis of Adenosine Triphosphate