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Rja Wanders - One of the best experts on this subject based on the ideXlab platform.
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Straight-chain Acyl-CoA Oxidase deficiency presenting with dysmorphia, neurodevelopmental autistic-type regression and a selective pattern of leukodystrophy
Journal of inherited metabolic disease, 2004Co-Authors: M. A. Kurian, Rja Wanders, Stephanie Ryan, G. T. N. Besley, Mary D. KingAbstract:Summary: We report a rare case of straight-chain Acyl-CoA Oxidase deficiency (McKusick 264470) presenting with dysmorphism, neurodevelopmental regression and leukodystrophy.
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Novel deletion in a patient with an isolated peroxisoml Acyl-CoA Oxidase deficiency
American Journal of Human Genetics, 1994Co-Authors: B. T. Poll-the, H. Clevers, B. Fournier, Rja WandersAbstract:Disorders with defective peroxisome assembly are associated with multiple peroxisomal enzymatic abnormalities. Besides these diseases patients have been described suspected of having a single enzyme defect in the peroxisomal {beta}-oxidation pathway. Laboratory findings for these patients include elevated plasma very long chain fatty acids (VLCFA) and impaired VLCFA oxidation in fibroblasts. Complementation analysis between these patients and those with a proven single enzyme deficiency, using peroxisomal {beta}-oxidation of VLCFA as the criterion for complementation, has been used to show whether the patients are deficient in Acyl-CoA Oxidase, peroxisomal trifunctional protein or thiolase activity. Fibroblasts from a patient showing the clinical and biochemical abnormalities of isolated Acyl-CoA Oxidase deficiency (using cell complementation) were analyzed at the molecular level. Isolation of RNA from patient`s fibroblasts was followed by random reverse transcription of RNA and PCR amplification. PCR products were blotted and hybridized with the human Acyl-CoA Oxidase cDNA. A fragment 150 bp shorter than normal was found. Upon sequencing, exon 7 was found to be deleted leading to a frameshift in the Acyl-CoA Oxidase mRNA. Southern blot analysis of the patient`s DNA did not reveal any deletion in contrast to two siblings previously reported as having a deletion of at least 17more » kb in the Acyl-CoA Oxidase gene.« less
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The CoA esters of 2-methyl-branched chain fatty acids and of the bile acid intermediates di- and trihydroxycoprostanic acids are oxidized by one single peroxisomal branched chain Acyl-CoA Oxidase in human liver and kidney.
The Journal of biological chemistry, 1993Co-Authors: G Vanhove, Rja Wanders, P. P. Van Veldhoven, H J Eyssen, Marc Fransen, Simone Denis, G P MannaertsAbstract:Rat liver peroxisomes contain three Acyl-CoA Oxidases: palmitoyl-CoA Oxidase, which oxidizes the CoA esters of straight chain fatty acids and prostaglandins; pristanoyl-CoA Oxidase, which oxidizes the CoA esters of 2-methyl-branched fatty acids (e.g. pristanic acid); and trihydroxycoprostanoyl-CoA Oxidase, which oxidizes the CoA esters of the bile acid intermediates di- and trihydroxycoprostanic acids (Van Veldhoven, P. P., Vanhove, G., Asselberghs, S., Eyssen, H. J., and Mannaerts, G. P. (1992) J. Biol. Chem. 267, 20065-20074). In the present report we demonstrate that human liver peroxisomes contain only two Acyl-CoA Oxidases: palmitoyl-CoA Oxidase, which oxidizes the CoA esters of straight chain fatty acids and prostaglandins, and a novel branched chain Acyl-CoA Oxidase, which oxidizes the CoA esters of 2-methyl-branched fatty acids as well as those of the bile acid intermediates (which also possess a 2-methyl substitution in their side chains). The branched chain Acyl-CoA Oxidase was purified to near homogeneity by means of column chromatography. It appeared to be a 70-kDa monomeric protein that did not cross-react with antisera raised against rat palmitoyl-CoA Oxidase and pristanoyl-CoA Oxidase. No indication was found for the presence of a separate trihydroxycoprostanoyl-CoA Oxidase in human liver. The branched chain Acyl-CoA Oxidase was present also in human kidney, suggesting that it is expressed in other extrahepatic tissues as well. Our results explain a number of clinical-chemical observations made in certain cases of peroxisomal beta-oxidation disorders.
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Peroxisomal beta-oxidation of polyunsaturated long chain fatty acids in human fibroblasts. The polyunsaturated and the saturated long chain fatty acids are retroconverted by the same Acyl-CoA Oxidase.
Scandinavian Journal of Clinical & Laboratory Investigation, 1993Co-Authors: Erik Christensen, Rja Wanders, B. T. Poll-the, Berit Woldseth, Tor-arne Hagve, H. Sprecher, Oddvar Stokke, Bjørn O. ChristophersenAbstract:The metabolism of the C22 unsaturated fatty acids erucic acid (22:1(n-9)), adrenic acid (22:4(n-6)), docosapentaenoic acid (22:5(n-3)) and docosahexaenoic acid (22:6(n-3)) was studied in cultured fibroblasts from patients with Acyl-CoA Oxidase deficiency, the Zellweger syndrome, X-linked adrenoleukodystrophy (X-ALD) and normal controls.[3-14C] 22:4 (n-6) and [3-14C] 22:5 (n-3) were shortened (retroconverted) to [1-I4C] 20:4 (n-6) and [1-14C] 20:5 (n-3), respectively, in normal and X-ALD fibroblasts. In Zellweger and Acyl-CoA Oxidase deficient fibroblasts these reactions were deficient.Since the retroconversion is normal in X-ALD fibroblasts peroxisomal very long chain (lignoceryl) CoA ligase is probably not required for the activation of C22 unsaturated fatty acids.The present work with fibroblasts from patients with a specific Acyl-CoA Oxidase deficiency, previously shown to have a deficient peroxisomal clofibrate-inducible Acyl-CoA Oxidase, and which accumulate 24:0 and 26:0 fatty acids, supports the vi...
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Peroxisomal β-oxidation of Polyunsaturated Long Chain Fatty Acids in Human Fibroblasts. The Polyunsaturated and the Saturated Long Chain Fatty Acids are Retroconverted by the Same Acyl-CoA Oxidase
Scandinavian Journal of Clinical and Laboratory Investigation, 1993Co-Authors: Erik Christensen, Rja Wanders, B. T. Poll-the, Berit Woldseth, Tor-arne Hagve, H. Sprecher, Oddvar Stokke, Bjørn O. ChristophersenAbstract:The metabolism of the C22 unsaturated fatty acids erucic acid (22:1(n-9)), adrenic acid (22:4(n-6)), docosapentaenoic acid (22:5(n-3)) and docosahexaenoic acid (22:6(n-3)) was studied in cultured fibroblasts from patients with Acyl-CoA Oxidase deficiency, the Zellweger syndrome, X-linked adrenoleukodystrophy (X-ALD) and normal controls. [3-14C] 22:4 (n-6) and [3-14C] 22:5 (n-3) were shortened (retroconverted) to [1-14C] 20:4 (n-6) and [1-14C] 20:5 (n-3), respectively, in normal and X-ALD fibroblasts. In Zellweger and Acyl-CoA Oxidase deficient fibroblasts these reactions were deficient. Since the retroconversion is normal in X-ALD fibroblasts peroxisomal very long chain (lignoceryl) CoA ligase is probably not required for the activation of C22 unsaturated fatty acids. The present work with fibroblasts from patients with a specific Acyl-CoA Oxidase deficiency, previously shown to have a deficient peroxisomal clofibrate-inducible Acyl-CoA Oxidase, and which accumulate 24:0 and 26:0 fatty acids, supports the view that this enzyme is responsible for the chain-shortening of docosahexaenoic acid (22:6(n-3)), erucic acid (22:1(n-9)), docosapentaenoic acid (22:5(n-3)), and adrenic acid (22:4(n-6)) as wel
Mikio Nishimura - One of the best experts on this subject based on the ideXlab platform.
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A Novel Acyl-CoA Oxidase That Can Oxidize Short-chain Acyl-CoA in Plant Peroxisomes
The Journal of biological chemistry, 1999Co-Authors: H. Hayashi, L. De Bellis, A. Ciurli, Maki Kondo, M. Hayashi, Mikio NishimuraAbstract:Abstract Short-chain Acyl-CoA Oxidases are β-oxidation enzymes that are active on short-chain Acyl-CoAs and that appear to be present in higher plant peroxisomes and absent in mammalian peroxisomes. Therefore, plant peroxisomes are capable of performing complete β-oxidation of Acyl-CoA chains, whereas mammalian peroxisomes can perform β-oxidation of only those Acyl-CoA chains that are larger than octanoyl-CoA (C8). In this report, we have shown that a novel Acyl-CoA Oxidase can oxidize short-chain Acyl-CoA in plant peroxisomes. A peroxisomal short-chain Acyl-CoA Oxidase from Arabidopsis was purified following the expression of the Arabidopsis cDNA in a baculovirus expression system. The purified enzyme was active on butyryl-CoA (C4), hexanoyl-CoA (C6), and octanoyl-CoA (C8). Cell fractionation and immunocytochemical analysis revealed that the short-chain Acyl-CoA Oxidase is localized in peroxisomes. The expression pattern of the short-chain Acyl-CoA Oxidase was similar to that of peroxisomal 3-ketoAcyl-CoA thiolase, a marker enzyme of fatty acid β-oxidation, during post-germinative growth. Although the molecular structure and amino acid sequence of the enzyme are similar to those of mammalian mitochondrial Acyl-CoA dehydrogenase, the purified enzyme has no activity as Acyl-CoA dehydrogenase. These results indicate that the short-chain Acyl-CoA Oxidases function in fatty acid β-oxidation in plant peroxisomes, and that by the cooperative action of long- and short-chain Acyl-CoA Oxidases, plant peroxisomes are capable of performing the complete β-oxidation of Acyl-CoA.
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Purification and characterization of pumpkin long‐chain acyl‐CoA Oxidase
Physiologia Plantarum, 1999Co-Authors: Luigi De Bellis, Pietro Giuntini, Hiroshi Hayashi, Makoto Hayashi, Mikio NishimuraAbstract:Pumpkin (Cucurbita sp.) long-chain Acyl-CoA Oxidase (ACOX) (EC 1.3.3.6) was purified to homogeneity by hydrophobic interaction, hydroxyapatite, affinity, and anion exchange chromatographies. The purified isoenzyme is a dimeric protein, consisting of two apparently identical 72-kDa subunits. The protein is exclusively localized in glyoxysomes. The enzyme catalyzes selectively the oxidation of CoA esters of fatty acids with 12-18 C atoms and exhibits highest activity with C-14 fatty acids, but no activity with isobutyryl-CoA and isovaleryl-CoA (branched chain) or glutaryl-CoA (dicarboxylic). The enzyme is strongly inhibited by high concentrations of palmitoyl-CoA and weakly inhibited by high concentration of myristoyl-CoA. It is also inhibited by Triton X-100 at concentrations above 0.018% (w/v) the critical micellar concentration. The consequences of the substrate inhibition for the evaluation of long-chain ACOX activity in plant tissues are discussed.
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Molecular characterization of a glyoxysomal long chain Acyl-CoA Oxidase that is synthesized as a precursor of higher molecular mass in pumpkin.
The Journal of biological chemistry, 1998Co-Authors: Hidenori Hayashi, L. De Bellis, M. Hayashi, Katsushi Yamaguchi, Akira Kato, Mikio NishimuraAbstract:Abstract A cDNA clone for pumpkin Acyl-CoA Oxidase (EC1.3.3.6; ACOX) was isolated from a λgt11 cDNA library constructed from poly(A)+ RNA extracted from etiolated cotyledons. The inserted cDNA clone contains 2313 nucleotides and encodes a polypeptide of 690 amino acids. Analysis of the amino-terminal sequence of the protein indicates that the pumpkin Acyl-CoA Oxidase protein is synthesized as a larger precursor containing a cleavable amino-terminal presequence of 45 amino acids. This presequence shows high similarity to the typical peroxisomal targeting signal (PTS2). Western blot analysis following cell fractionation in a sucrose gradient revealed that ACOX is localized in glyoxysomes. A partial purification of ACOX from etiolated pumpkin cotyledons indicated that the ACOX cDNA codes for a long chain Acyl-CoA Oxidase. The amount of ACOX increased and reached to the maximum activity by day 5 of germination but decreased about 4-fold on the following days during the subsequent microbody transition from glyoxysomes to leaf peroxisomes. By contrast, the amount of mRNA was already high at day 1 of germination, increased by about 30% at day 3, and faded completely by day 7. These data indicated that the expression pattern of ACOX was very similar to that of the glyoxysomal enzyme 3-ketoAcyl-CoA thiolase, another marker enzyme of the β-oxidation spiral, during germination and suggested that the expression of each enzyme of β-oxidation is coordinately regulated.
Bjørn O. Christophersen - One of the best experts on this subject based on the ideXlab platform.
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Peroxisomal beta-oxidation of polyunsaturated long chain fatty acids in human fibroblasts. The polyunsaturated and the saturated long chain fatty acids are retroconverted by the same Acyl-CoA Oxidase.
Scandinavian Journal of Clinical & Laboratory Investigation, 1993Co-Authors: Erik Christensen, Rja Wanders, B. T. Poll-the, Berit Woldseth, Tor-arne Hagve, H. Sprecher, Oddvar Stokke, Bjørn O. ChristophersenAbstract:The metabolism of the C22 unsaturated fatty acids erucic acid (22:1(n-9)), adrenic acid (22:4(n-6)), docosapentaenoic acid (22:5(n-3)) and docosahexaenoic acid (22:6(n-3)) was studied in cultured fibroblasts from patients with Acyl-CoA Oxidase deficiency, the Zellweger syndrome, X-linked adrenoleukodystrophy (X-ALD) and normal controls.[3-14C] 22:4 (n-6) and [3-14C] 22:5 (n-3) were shortened (retroconverted) to [1-I4C] 20:4 (n-6) and [1-14C] 20:5 (n-3), respectively, in normal and X-ALD fibroblasts. In Zellweger and Acyl-CoA Oxidase deficient fibroblasts these reactions were deficient.Since the retroconversion is normal in X-ALD fibroblasts peroxisomal very long chain (lignoceryl) CoA ligase is probably not required for the activation of C22 unsaturated fatty acids.The present work with fibroblasts from patients with a specific Acyl-CoA Oxidase deficiency, previously shown to have a deficient peroxisomal clofibrate-inducible Acyl-CoA Oxidase, and which accumulate 24:0 and 26:0 fatty acids, supports the vi...
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Peroxisomal β-oxidation of Polyunsaturated Long Chain Fatty Acids in Human Fibroblasts. The Polyunsaturated and the Saturated Long Chain Fatty Acids are Retroconverted by the Same Acyl-CoA Oxidase
Scandinavian Journal of Clinical and Laboratory Investigation, 1993Co-Authors: Erik Christensen, Rja Wanders, B. T. Poll-the, Berit Woldseth, Tor-arne Hagve, H. Sprecher, Oddvar Stokke, Bjørn O. ChristophersenAbstract:The metabolism of the C22 unsaturated fatty acids erucic acid (22:1(n-9)), adrenic acid (22:4(n-6)), docosapentaenoic acid (22:5(n-3)) and docosahexaenoic acid (22:6(n-3)) was studied in cultured fibroblasts from patients with Acyl-CoA Oxidase deficiency, the Zellweger syndrome, X-linked adrenoleukodystrophy (X-ALD) and normal controls. [3-14C] 22:4 (n-6) and [3-14C] 22:5 (n-3) were shortened (retroconverted) to [1-14C] 20:4 (n-6) and [1-14C] 20:5 (n-3), respectively, in normal and X-ALD fibroblasts. In Zellweger and Acyl-CoA Oxidase deficient fibroblasts these reactions were deficient. Since the retroconversion is normal in X-ALD fibroblasts peroxisomal very long chain (lignoceryl) CoA ligase is probably not required for the activation of C22 unsaturated fatty acids. The present work with fibroblasts from patients with a specific Acyl-CoA Oxidase deficiency, previously shown to have a deficient peroxisomal clofibrate-inducible Acyl-CoA Oxidase, and which accumulate 24:0 and 26:0 fatty acids, supports the view that this enzyme is responsible for the chain-shortening of docosahexaenoic acid (22:6(n-3)), erucic acid (22:1(n-9)), docosapentaenoic acid (22:5(n-3)), and adrenic acid (22:4(n-6)) as wel
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Peroxisomal beta-oxidation of polyunsaturated long chain fatty acids in human fibroblasts. The polyunsaturated and the saturated long chain fatty acids are retroconverted by the same Acyl-CoA Oxidase.
Scandinavian journal of clinical and laboratory investigation. Supplementum, 1993Co-Authors: Erik Christensen, B. T. Poll-the, Berit Woldseth, Tor-arne Hagve, H. Sprecher, Oddvar Stokke, R J Wanders, Bjørn O. ChristophersenAbstract:The metabolism of the C22 unsaturated fatty acids erucic acid (22:1(n-9)), adrenic acid (22:4(n-6)), docosapentaenoic acid (22:5(n-3)) and docosahexaenoic acid (22:6(n-3)) was studied in cultured fibroblasts from patients with Acyl-CoA Oxidase deficiency, the Zellweger syndrome, X-linked adrenoleukodystrophy (X-ALD) and normal controls. [3-14C] 22:4 (n-6) and [3-14C] 22:5 (n-3) were shortened (retroconverted) to [1-14C] 20:4 (n-6) and [1-14C] 20:5 (n-3), respectively, in normal and X-ALD fibroblasts. In Zellweger and Acyl-CoA Oxidase deficient fibroblasts these reactions were deficient. Since the retroconversion is normal in X-ALD fibroblasts peroxisomal very long chain (lignoceryl) CoA ligase is probably not required for the activation of C22 unsaturated fatty acids. The present work with fibroblasts from patients with a specific Acyl-CoA Oxidase deficiency, previously shown to have a deficient peroxisomal clofibrate-inducible Acyl-CoA Oxidase, and which accumulate 24:0 and 26:0 fatty acids, supports the view that this enzyme is responsible for the chain-shortening of docosahexaenoic acid (22:6(n-3)), erucic acid (22:1(n-9)), docosapentaenoic acid (22:5(n-3)), and adrenic acid (22:4(n-6)) as well.
Clifford R. Elcombe - One of the best experts on this subject based on the ideXlab platform.
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Differential tissue‐specific expression and induction of cytochrome P450IVA1 and acyl‐CoA Oxidase
European journal of biochemistry, 1992Co-Authors: David R. Bell, Remi Bars, Clifford R. ElcombeAbstract:We have examined the tissue-specific expression and inducibility of Acyl-CoA Oxidase and cytochrome P450IVA1 (P450IVA1) RNA in rats. Groups of three rats were dosed daily by gavage with methylclofenapate at 25 mg/kg in 5 ml/kg corn oil for nine weeks, or were administered a vehicle control. P450IVA1 and Acyl-CoA Oxidase RNA were detected using an RNase protection assay. Similar levels of Acyl-CoA Oxidase RNA were present in control liver and kidney, but the level of this RNA in lung, muscle and testis was 6–11%, and in pancreas was 0.13%, of that in liver. Treatment of rats with methylclofenapate led to an 11-fold induction of Acyl-CoA Oxidase RNA in liver and also produced a significant induction of this RNA in kidney, lung, muscle and testis of 1.7-fold, 1.3-fold, 2-fold and 1.7-fold, respectively. Acyl-CoA Oxidase RNA was not induced in pancreas. P450IVA1 RNA was present in control liver and also in kidney of control rats at 28% of the level in liver. In contrast to Acyl-CoA Oxidase RNA, P450IVA1 RNA was not detected in lung, pancreas or testis. Methylclofenapate treatment of rats led to an 18-fold induction of P450IVA1 RNA in liver, and a sevenfold induction in kidney. Induction of P450IVA1 was not detected in any of the other tissues examined. Quantification of the relative amounts of Acyl-CoA Oxidase and P450IVA1 RNA in control liver revealed that Acyl-CoA Oxidase RNA was present in a 17.5-fold molar excess over P450IVA1 RNA. Western blotting with an anti-P450IVA IgG revealed two bands of similar apparent molecular mass in liver and kidney microsomes, but not in microsomes from the testis of control rats. Methylclofenapate treatment of rats caused an increase in the intensity of these bands in microsomes from liver, but no induction was obvious in kidney. Immunocytochemical staining for both the microsomal P450IVA and peroxisomal Acyl-CoA Oxidase proteins was restricted to the proximal convoluted tubule in the kidney cortex, with staining being most intense in the S3 region.
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Regulation of differentially spliced transcripts of Acyl-CoA Oxidase in the rat.
Biochimica et biophysica acta, 1991Co-Authors: David R. Bell, Clifford R. ElcombeAbstract:RNAse protection assay was used to distinguish between and to quantify alternatively spliced transcripts of Acyl-CoA Oxidase in liver, kidney and testis of control and methylclofenapate treated rats. The ratio of spliced transcripts (type I to II) was 1.18:1 in control liver RNA, with 130 and 110 molecules/cell, respectively, and 3.1:1 in treated liver RNA, with 2800 and 900 molecules/cell. The ratios were 1.6 and 2:1 in control and treated kidney, and 0.31:1 in testis. This is likely to be due to differential splicing, which is, therefore, regulated during peroxisome proliferation, and also in a tissue specific fashion.
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Localization and differential induction of cytochrome P450IVA and Acyl-CoA Oxidase in rat liver
Biochemical Journal, 1991Co-Authors: David R. Bell, Remi Bars, G. Gordon Gibson, Clifford R. ElcombeAbstract:The peroxisome proliferators are structurally diverse chemicals which induce hyperplasia, hypertrophy and the proliferation of peroxisomes in the rodent liver. Cytochrome P450IVA1 and peroxisomal enzymes, such as Acyl-CoA Oxidase, are induced and are early markers of treatment with peroxisome proliferators. In this study, rats were dosed intraperitoneally with the potent peroxisome proliferator methylclofenapate and the hepatic induction response was studied. There was no significant change in the enzyme activities of laurate hydroxylase (cytochrome P450IVA1) or Acyl-CoA Oxidase in the first 8 h after treatment, but the activities had doubled at 24 h, suggesting that these enzymes are not involved in the mediation of early events in peroxisome proliferation. Hepatic cytochrome P450IVA1 mRNA was significantly increased at 6 and 8 h after treatment, rising to 15-fold above control values at 30 h. In contrast, Acyl-CoA Oxidase mRNA showed no significant change in the first 8 h, but increased to 13-fold above control values at 24 and 30 h, thereby demonstrating different kinetics of induction of the two mRNAs. In order to determine whether cytochrome P450IVA1 and peroxisomal enzymes were included in the same cells, rats were treated daily with sub-maximal (2 or 5 mg/kg) and maximal (25 mg/kg) inducing doses of methylclofenapate for 4 days. The lobular distribution of induced proteins was determined immunocytochemically with antibodies raised against P450IVA1 and Acyl-CoA Oxidase. Livers from control animals showed minimal staining for both proteins. However, in the livers of animals treated with 2 or 5 mg of methylclofenapate/kg, both Acyl-CoA and P450IVA immunostaining was increased, mainly in the centrilobular area. Immunostaining of serial sections revealed that these proteins were induced in the same region of the lobule. A maximal inducing dose of methylclofenapate (25 mg/kg) caused panlobular induction of both proteins. The results demonstrate that these proteins are induced in a dose-dependent manner in the same, spatially distinct, sensitive region of the liver lobule.
Retsu Miura - One of the best experts on this subject based on the ideXlab platform.
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Crystallization and preliminary X-ray characterization of rat liver Acyl-CoA Oxidase
Acta Crystallographica Section D Biological Crystallography, 2001Co-Authors: Yoshitaka Nakajima, Chiaki Setoyama, Haruhiko Tamaoki, Yasuzo Nishina, Kiyoshi Shiga, Ikuko Miyahara, Ken Hirotsu, Retsu MiuraAbstract:A recombinant form of the flavoenzyme Acyl-CoA Oxidase from rat liver has been crystallized by the hanging-drop vapour-diffusion technique using PEG 20 000 as a precipitating agent. The crystals grew as yellow prisms, with unit-cell parameters a = 71.05, b = 87.29, c = 213.05 A, α = β = γ = 90°. The crystals exhibit the symmetry of space group P212121 and are most likely to contain a dimer in the asymmetric unit, with a VM value of 2.21 A3 Da−1. The crystals diffract to a resolution of 2.5 A at beamline BL6A of the Photon Factory. Two heavy-atom derivatives have been identified.
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Functional expression of two forms of rat Acyl-CoA Oxidase and their substrate specificities.
Biochemical and biophysical research communications, 1995Co-Authors: Chiaki Setoyama, Haruhiko Tamaoki, Yasuzo Nishina, Kiyoshi Shiga, Retsu MiuraAbstract:Using the reverse transcription of RNA followed by the polymerase chain reaction, we cloned the cDNAs for the rat Acyl-CoA Oxidases I and II, which are produced by alternative splicing from a single gene, and developed a system for their expression in Escherichia coli. The homogeneous preparations of these enzymes, without proteolytic procession, showed Oxidase activity with Acyl-CoAs having various acyl-chain lengths. The two types of the enzyme exhibited different substrate specificities with respect to the acyl-chain length, Acyl-CoA Oxidase I showing the optimum activity at shorter chain-length relative to Acyl-CoA Oxidase II.
