3-Hydroxyacyl-CoA Dehydrogenase

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Takashi Hashimoto - One of the best experts on this subject based on the ideXlab platform.

  • Amino acid and nucleotide sequences of human peroxisomal enoyl-CoA hydratase : 3-Hydroxyacyl-CoA Dehydrogenase cDNA
    Journal of inherited metabolic disease, 1998
    Co-Authors: Seiji Fukuda, Tadao Orii, Yasuyuki Suzuki, Nobuyuki Shimozawa, Zhongyi Zhang, Toshifumi Aoyama, Takashi Hashimoto, Naomi Kondo
    Abstract:

    Deficiency of enoyl-CoA hydratase : 3-Hydroxyacyl-CoA Dehydrogenase (peroxisomal bifunctional enzyme), one of the enzymes of the peroxisomal β-oxidation system, leads to clinical manifestations resembling Zellweger syndrome with hypotonia, psychomotor delay, hepatomegaly, typical facial appearance and accumulation of very long-chain fatty acids. The nucleotide sequence of the human peroxisomal enoyl-CoA hydratase : 3-Hydroxyacyl-CoA Dehydrogenase cDNA has been reported by Hoefler and colleagues; however, we have found some amino acid differences from our originally isolated cDNA. Contrary to the findings described in a previous paper, we report here the cDNA sequence of human peroxisomal enoyl-CoA hydratase : 3-Hydroxyacyl-CoA Dehydrogenase in which there are 9 authenticated amino acid alterations.

  • Physiological Role of D-3-Hydroxyacyl-CoA Dehydratase/D-3-Hydroxyacyl-CoA Dehydrogenase Bifunctional Protein.
    Journal of biochemistry, 1997
    Co-Authors: Ling Ling Jiang, Yasuyuki Suzuki, Takao Kurosawa, Masahiro Sato, Takashi Hashimoto
    Abstract:

    The second and third reactions of the peroxisomal beta-oxidation spiral are thought to be catalyzed by enoyl-CoA hydratase/L-3-Hydroxyacyl-CoA Dehydrogenase bifunctional protein (L-bifunctional protein). Recently, we found the presence of D-3-Hydroxyacyl-CoA dehydratase/D-3-Hydroxyacyl-CoA Dehydrogenase bifunctional protein (D-bifunctional protein) in mammalian peroxisomes. Therefore, we studied the physiological role of the D-bifunctional protein. The contents of the L- and D-bifunctional proteins were about 0.01 and 0.5 microg/mg protein, respectively, in cultured human skin fibroblasts. The activity of conversion of hexadecenoyl-CoA to 3-ketopalmitoyl-CoA by the D-bifunctional protein was estimated to be about 0.5 milliunit/mg of fibroblast protein. This value was about 100-fold that of the L-bifunctional protein in the fibroblasts. From comparison of the activities of the bifunctional proteins with the rate of palmitate oxidation and the activities of acyl-CoA oxidase and 3-ketoacyl-CoA thiolase, it is proposed that the D-bifunctional protein plays a major role in the peroxisomal oxidation of palmitate in the fibroblasts. The contents of both the L- and D-bifunctional proteins in liver were about 2.5 microg/mg protein. Therefore, it is suggested that the D-bifunctional protein also plays a significant role in human liver peroxisomal fatty acid oxidation. Actions of the bifunctional proteins on enoyl forms of other acyl-CoA derivatives were examined. The D-bifunctional protein but not the L-bifunctional protein reacted with 2-methylhexadecenoyl-CoA and 3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-2 4-enoyl-CoA. We propose that, among the reactions of the distinct group of carboxylates oxidized specifically in peroxisomes, oxidation of 2-methyl-branched fatty acids and side-chain shortening of cholesterol for bile acid formation are catalyzed by the D-bifunctional protein, but not the L-bifunctional protein.

  • Cloning and expression of cDNA for a newly identified isozyme of bovine liver 3-Hydroxyacyl-CoA Dehydrogenase and its import into mitochondria.
    Biochimica et biophysica acta, 1997
    Co-Authors: Shuichi Furuta, Shoko Miyazawa, Akio Kobayashi, Takashi Hashimoto
    Abstract:

    cDNA for a heretofore undescribed mitochondrial 3-Hydroxyacyl-CoA Dehydrogenase, designated as the type II enzyme with different molecular and catalytic properties, compared to those of the classical mitochondrial beta-oxidation enzyme (type I enzyme), was cloned from a bovine liver cDNA library. Nucleotide sequence of the cDNA encoded 261 amino acids with a subunit molecular weight of 27,140. The deduced primary structure of the type II enzyme showed no significant homology to the reported amino acid sequence of the classical 3-Hydroxyacyl-CoA Dehydrogenases. On SDS-PAGE, no differences in subunit molecular weights were observed among the in vitro translation products, the recombinant type II enzyme produced in Escherichia coli and the purified enzyme. NH2-terminal and COOH-terminal amino acid sequence analysis of the purified type II enzyme revealed that the mature enzyme had not been proteolytically processed. The in vitro translation products of the type II enzyme were efficiently incorporated into isolated rat liver mitochondria, without changes in size, thereby suggesting that unlike other mitochondrial enzymes of fatty acid beta-oxidation, the type II enzyme had no cleavable signal peptide upon import into mitochondria.

  • Structure of D-3-Hydroxyacyl-CoA Dehydratase/D-3-Hydroxyacyl-CoA Dehydrogenase Bifunctional Protein
    Journal of biochemistry, 1997
    Co-Authors: Ling Ling Jiang, Masayoshi Souri, Shoko Miyazawa, Takashi Hashimoto
    Abstract:

    When D-3-Hydroxyacyl-CoA dehydratase/D-3-Hydroxyacyl-CoA Dehydrogenase bifunctional protein was purified from human liver, two preparations were obtained. One contained a 77-kDa polypeptides as the main and minor smaller polypeptides including a 46-kDa polypeptide, and this preparation showed both the dehydratase and Dehydrogenase activities. The other preparation was a homodimer of the 46-kDa polypeptide and showed only the dehydratase activity. Further analysis indicated that the native enzyme is a homodimer of 77-kDa polypeptide, which was proteolytically modified during purification. The cDNA for the human 77-kDa polypeptide was cloned. The amino acid sequences of the peptides derived from the components of the enzyme preparations were located in the deduced amino acid sequence of the cDNA. The preparation containing the 77-kDa polypeptide was treated with a protease, and two monofunctional fragments were separated. The Dehydrogenase and dehydratase fragments were located on the amino- and carboxyl-terminal sides, respectively, of the deduced amino acid sequence of the cDNA. The protein expressed by the cDNA with the entire coding region exhibited both the dehydratase and Dehydrogenase activities, and that expressed by a truncated version covering the carboxyl-terminal side exhibited only the dehydratase activity. The cloned cDNA was identical to the human 17 beta-hydroxysteroid Dehydrogenase IV cDNA.

  • Purification and Properties of Rat D-3-Hydroxyacyl-CoA Dehydratase: D-3-Hydroxyacyl-CoA Dehydratase/D-3-Hydroxyacyl-CoA Dehydrogenase Bifunctional Protein
    Journal of biochemistry, 1996
    Co-Authors: Ling Ling Jiang, Shoko Miyazawa, Takashi Hashimoto
    Abstract:

    We have previously purified two D-3-Hydroxyacyl-CoA dehydratase preparations from human liver. One preparation contained a 77-kDa polypeptide and smaller polypeptides, and the other was a homodimer of a 46-kDa polypeptide. Three different purified rat peroxisomal D-3-Hydroxyacyl-CoA dehydratase preparations have been reported. Therefore, rat enzyme was purified in this study to confirm the enzyme structure. Two preparations with similar molecular structures to the human enzyme preparations were obtained, and these were similar to each other in immunochemical and catalytic properties. It was suggested that the native enzyme was a homodimer of the 77-kDa polypeptide, and this enzyme was modified to a homodimer of the 46-kDa polypeptide, because conversion of the 77-kDa polypeptide to smaller polypeptides including the 46-kDa polypeptide was clearly observed during purification. Rat liver subcellular fractionation study indicates that this enzyme is located in peroxisomes. The enzyme preparation containing the 77-kDa polypeptide catalyzed the D-3-Hydroxyacyl-CoA Dehydrogenase reaction as well as the dehydratase reaction. Thus, it is proposed that this enzyme is D-3-Hydroxyacyl-CoA dehydratase/ D-3-Hydroxyacyl-CoA Dehydrogenase bifunctional protein.

R. J. A. Wanders - One of the best experts on this subject based on the ideXlab platform.

  • Striking improvement of muscle strength under creatine therapy in a patient with long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency.
    Journal of inherited metabolic disease, 2003
    Co-Authors: G. C. Korenke, R. J. A. Wanders, Folker Hanefeld
    Abstract:

    Summary: Creatine monohydrate given orally led to a long-lasting improvement of muscular weakness and ataxia in a girl with long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency

  • Secondary respiratory chain defect in a boy with long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency: possible diagnostic pitfalls
    European journal of pediatrics, 2000
    Co-Authors: A. M. Das, R. J. A. Wanders, R. Fingerhut, Kurt Ullrich
    Abstract:

    We report on a boy who suffered from microcephaly, growth retardation, cardiomyopathy and hepatic dysfunction. When he had his first febrile infection at the age of 3 months he showed metabolic decompensation. Laboratory parameters and clinical features were compatible with a β-oxidation defect or a respiratory chain disorder. Measurement of β-oxidation enzymes showed long-chain 3-hydroxyacyl CoA Dehydrogenase (LCHAD) deficiency; determination of respiratory chain complex activities revealed complete absence of complex I, II, III and IV activities in skeletal muscle and reduced activities of complexes II and IV in cultured fibroblasts, with secondary dysregulation of ATP synthase. The patient was found to be homozygous for the MTP:G1528 C mutation (LCHAD-deficiency).

  • Molecular basis of long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency: identification of two new mutations.
    Journal of inherited metabolic disease, 1997
    Co-Authors: Lodewijk Ijlst, Jos P.n. Ruiter, W. Oostheim, R. J. A. Wanders
    Abstract:

    Long-chain 3-Hydroxyacyl-CoA Dehydrogenase (LCHAD) is catalysed by the mitochondrial trifunctional protein (MTP), which also contains enoyl-CoA hydratase and 3-ketothiolase activities (Carpenter et al 1992; Uchida et al 1992). The cDNAs encoding the a and β subunits were cloned by Kamijo et al (1994a). Many patients have been described with a defect in this enzyme complex and it appears that in most patients there is an isolated deficiency of the Dehydrogenase activity of the MTP. We and others have reported a G1528C mutation in the gene coding for the α subunit of MTP, changing the codon for glutamate (510) into glutamine (IJ1st et al 1994; Sims et al 1995). In a series of 34 LCHAD-deficient patients the G1528C mutation was found to be very frequent (87%), which corresponds to the situation observed in MCAD deficiency with the frequent G985A mutation. The G1528C mutation is directly responsible for the loss of Dehydrogenase activity without changing the structure of the enzyme complex (IJ1st et al 1996). In a group of 46 LCHAD-deficient patients as studied enzymatically in our laboratory, we found 12 to be compound heterozygous for the common mutation. Here we describe two new mutations found in this compound heterozygous group.

  • Long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency: A new method to identify the G1528C mutation in genomic DNA showing its high frequency (≈90%) and identification of a new mutation (T2198C)
    Journal of inherited metabolic disease, 1996
    Co-Authors: L. Ijlst, Jos P.n. Ruiter, J. Vreijling, R. J. A. Wanders
    Abstract:

    Long-chain 3-Hydroxyacyl-CoA Dehydrogenase (LC3HAD) is not a monofunctional enzyme like all the other enzymes involved in mitochondrial β-oxidation but instead is part of a trifunctional enzyme also harbouring long-chain enoyl-CoA hydratase and long-chain 3-ketothiolase activity. Purification of the enzyme protein was reported in 1992 by two groups of investigators (Carpenter et al 1992 ; Uchida et al 1992) and it appears to be an octamer of four α- and four β-subunits. Since its first description in 1989, many cases of long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency have been described. Biochemically, two phenotypes can be distinguished : in one group of patients (I), mitochondrial trifunctional protein (MTP) is completely deficient, as reflected in the absence of both the α- and the β-subunits upon immunoblotting. When measured in total homogenates prepared from fibroblasts from MTP-deficient patients, long-chain enoyl-CoA hydratase, long-chain 3-Hydroxyacyl-CoA Dehydrogenase and long-chain thiolase are all deficient, although to different extents (43%, 25% and 1.7% of control values, respectively) (Ijlst et al 1994). In the second group of LC3HAD-deficient patients (II), the enzyme protein is present, as shown by immunoblotting, although the level is somewhat lower than in control fibroblasts. This explains why as well as long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency (25% of mean control) there is also a partial deficiency of long-chain enoyl-CoA hydratase and long-chain 3-ketothiolase with residual activities of 78% and 59%, respectively (Ijlst et al 1994). We recently reported the identification of a point mutation at position 1528 of the cDNA for the α-subunit of mitochondrial trifunctional protein in fibroblasts from LC3HAD-deficient patients of group II. In these studies it was found that the G1528C mutation is frequent among LC3HAD-deficient patients. The exact frequency, however, could not be established because mutation analysis could only be done at the cDNA level. We have now set up a method using genomic DNA and have used it to determine the true frequency of the G1528C mutation. The results obtained are reported here. We also describe the identification of a new mutation (T2198C) in a patient heterozygous for the G1528C mutation.

  • A simple, straightforward assay for long-chain 3-Hydroxyacyl-CoA Dehydrogenase based on the use of N-ethylmaleimide: potential for pre- and postnatal diagnosis.
    Journal of inherited metabolic disease, 1993
    Co-Authors: L. Ijlst, R. J. A. Wanders
    Abstract:

    In recent years an increasing number of inherited diseases in man have been identified in which there is an impairment in mitochondrial fatty acid β-oxidation. This includes long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency, first identified in 1989 (Wanders et al 1989) and now described in the literature in at least twelve additional patients. Identification of long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency in patients suspected to suffer from this enzyme defect is usually done in cultured skin fibroblasts

Khalid Hussain - One of the best experts on this subject based on the ideXlab platform.

  • novel insights into fatty acid oxidation amino acid metabolism and insulin secretion from studying patients with loss of function mutations in 3 hydroxyacyl coa Dehydrogenase
    The Journal of Clinical Endocrinology and Metabolism, 2013
    Co-Authors: Amanda Heslegrave, Khalid Hussain
    Abstract:

    Context: Mutations causing genetic defects have been described in many of the enzymes involved in mitochondrial fatty acid oxidation (FAO). Recently, mutations in the penultimate enzyme in the FAO chain have been described that result in quite different symptoms from those normally seen. Patients with mutations in 3-Hydroxyacyl-CoA Dehydrogenase (HADH) present with protein (leucine)-induced hyperinsulinemic hypoglycemia (HH), suggesting a link between FAO, amino acid metabolism, and insulin secretion. Evidence Acquisition and Synthesis: Peer-reviewed articles were searched in PubMed with relevance to HADH and disorders of FAO and protein sensitivity. Relevant articles were cited. Recent evidence suggests that mutations in HADH cause HH that is precipitated by protein in a similar manner to the hyperinsulinism/hyperammonemia (HI/HA) syndrome, which is caused by mutations in the GLUD1 gene, encoding the enzyme glutamate Dehydrogenase (GDH). Conclusion: Current data suggest that the HH observed in patients w...

  • Leucine-sensitive hyperinsulinaemic hypoglycaemia in patients with loss of function mutations in 3-Hydroxyacyl-CoA Dehydrogenase.
    Orphanet Journal of Rare Diseases, 2012
    Co-Authors: Amanda Heslegrave, Simon Eaton, Ritika R. Kapoor, B. Chadefaux, Teoman Akcay, Enver Simsek, Sarah E. Flanagan, Sian Ellard, Khalid Hussain
    Abstract:

    Background: Loss of function mutations in 3-Hydroxyacyl-CoA Dehydrogenase (HADH) cause protein sensitive hyperinsulinaemic hypoglycaemia (HH). HADH encodes short chain 3-hydroxacyl-CoA Dehydrogenase, an enzyme that catalyses the penultimate reaction in mitochondrial β-oxidation of fatty acids. Mutations in GLUD1 encoding glutamate Dehydrogenase, also cause protein sensitive HH (due to leucine sensitivity). Reports suggest a protein-protein interaction between HADH and GDH. This study was undertaken in order to understand the mechanism of protein sensitivity in patients with HADH mutations. Methods: An oral leucine tolerance test was conducted in controls and nine patients with HADH mutations. Basal GDH activity and the effect of GTP were determined in lymphoblast homogenates from 4 patients and 3 controls. Immunoprecipitation was conducted in patient and control lymphoblasts to investigate protein interactions. Results: Patients demonstrated severe HH (glucose range 1.7–3.2 mmol/l; insulin range 4.8-63.8 mU/l) in response to the oral leucine load, this HH was not observed in control patients subjected to the same leucine load. Basal GDH activity and half maximal inhibitory concentration of GTP was similar in patients and controls. HADH protein could be co-immunoprecipitated with GDH protein in control samples but not in patient samples. Conclusions: We conclude that GDH and HADH have a direct protein-protein interaction, which is lost in patients with HADH mutations causing leucine induced HH. This is not associated with loss of inhibitory effect of GTP on GDH (as in patients with GLUD1 mutations).

  • Short-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency associated with hyperinsulinism: a novel glucose-fatty acid cycle?
    Biochemical Society Transactions, 2003
    Co-Authors: Simon Eaton, I. Chatziandreou, Steve Krywawych, S. Pen, Peter E. Clayton, Khalid Hussain
    Abstract:

    Hyperinsulinism of infancy is caused by inappropriate insulin secretion in pancreatic beta-cells, even when blood glucose is low. Several molecular defects are known to cause hyperinsulinism of infancy, such as K(ATP) channelopathies and regulatory defects of glucokinase and glutamate Dehydrogenase. Although defects of fatty acid oxidation have not previously been known to cause hyperinsulinism, patients with deficiency in SCHAD (short-chain 3-Hydroxyacyl-CoA Dehydrogenase; an enzyme of mitochondrial beta-oxidation) have hyperinsulinism. A novel link between fatty acid oxidation and insulin secretion may explain hyperinsulinism in these patients.

  • Hyperinsulinism in short-chain L-3-Hydroxyacyl-CoA Dehydrogenase deficiency reveals the importance of β-oxidation in insulin secretion
    The Journal of clinical investigation, 2001
    Co-Authors: Peter E. Clayton, Khalid Hussain, Simon Eaton, Steve Krywawych, Albert Aynsley-green, Mark Edginton, Vipan Datta, Helga E.m. Malingré, Ruud Berger, Inge E.t. Van Den Berg
    Abstract:

    A female infant of nonconsanguineous Indian parents presented at 4 months with a hypoglycemic convulsion. Further episodes of hypoketotic hypoglycemia were associated with inappropriately elevated plasma insulin concentrations. However, unlike other children with hyperinsulinism, this patient had a persistently elevated blood spot hydroxybutyrylcarnitine concentration when fed, as well as when fasted. Measurement of the activity of L-3-Hydroxyacyl-CoA Dehydrogenase in cultured skin fibroblasts with acetoacetyl-CoA substrate showed reduced activity. In fibroblast mitochondria, the activity was less than 5% that of controls. Sequencing of the short-chain L-3-Hydroxyacyl-CoA Dehydrogenase (SCHAD) genomic DNA from the fibroblasts showed a homozygous mutation (C773T) changing proline to leucine at amino acid 258. Analysis of blood from the parents showed they were heterozygous for this mutation. Western blot studies showed undetectable levels of immunoreactive SCHAD protein in the child's fibroblasts. Expression studies showed that the P258L enzyme had no catalytic activity. We conclude that C773T is a disease-causing SCHAD mutation. This is the first defect in fatty acid beta-oxidation that has been associated with hyperinsulinism and raises interesting questions about the ways in which changes in fatty acid and ketone body metabolism modulate insulin secretion by the beta cell. The patient's hyperinsulinism was easily controlled with diazoxide and chlorothiazide.

L. Ijlst - One of the best experts on this subject based on the ideXlab platform.

  • Long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency: A new method to identify the G1528C mutation in genomic DNA showing its high frequency (≈90%) and identification of a new mutation (T2198C)
    Journal of inherited metabolic disease, 1996
    Co-Authors: L. Ijlst, Jos P.n. Ruiter, J. Vreijling, R. J. A. Wanders
    Abstract:

    Long-chain 3-Hydroxyacyl-CoA Dehydrogenase (LC3HAD) is not a monofunctional enzyme like all the other enzymes involved in mitochondrial β-oxidation but instead is part of a trifunctional enzyme also harbouring long-chain enoyl-CoA hydratase and long-chain 3-ketothiolase activity. Purification of the enzyme protein was reported in 1992 by two groups of investigators (Carpenter et al 1992 ; Uchida et al 1992) and it appears to be an octamer of four α- and four β-subunits. Since its first description in 1989, many cases of long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency have been described. Biochemically, two phenotypes can be distinguished : in one group of patients (I), mitochondrial trifunctional protein (MTP) is completely deficient, as reflected in the absence of both the α- and the β-subunits upon immunoblotting. When measured in total homogenates prepared from fibroblasts from MTP-deficient patients, long-chain enoyl-CoA hydratase, long-chain 3-Hydroxyacyl-CoA Dehydrogenase and long-chain thiolase are all deficient, although to different extents (43%, 25% and 1.7% of control values, respectively) (Ijlst et al 1994). In the second group of LC3HAD-deficient patients (II), the enzyme protein is present, as shown by immunoblotting, although the level is somewhat lower than in control fibroblasts. This explains why as well as long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency (25% of mean control) there is also a partial deficiency of long-chain enoyl-CoA hydratase and long-chain 3-ketothiolase with residual activities of 78% and 59%, respectively (Ijlst et al 1994). We recently reported the identification of a point mutation at position 1528 of the cDNA for the α-subunit of mitochondrial trifunctional protein in fibroblasts from LC3HAD-deficient patients of group II. In these studies it was found that the G1528C mutation is frequent among LC3HAD-deficient patients. The exact frequency, however, could not be established because mutation analysis could only be done at the cDNA level. We have now set up a method using genomic DNA and have used it to determine the true frequency of the G1528C mutation. The results obtained are reported here. We also describe the identification of a new mutation (T2198C) in a patient heterozygous for the G1528C mutation.

  • First report of prenatal diagnosis of long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency in a pregnancy at risk.
    Prenatal diagnosis, 1993
    Co-Authors: Celia Pérez-cerdá, L. Ijlst, Ronald J.a. Wanders, Begoña Merinero, A. Jiménez, M. J. García, P. Sanz, Magdalena Ugarte
    Abstract:

    Prenatal diagnosis of long-chain 3-Hydroxyacyl-CoA Dehydrogenase (3-HAD) deficiency was performed in a family at risk. The diagnosis of an affected fetus was carried out by enzyme assay in cultured chorionic villus cells.

  • A simple, straightforward assay for long-chain 3-Hydroxyacyl-CoA Dehydrogenase based on the use of N-ethylmaleimide: potential for pre- and postnatal diagnosis.
    Journal of inherited metabolic disease, 1993
    Co-Authors: L. Ijlst, R. J. A. Wanders
    Abstract:

    In recent years an increasing number of inherited diseases in man have been identified in which there is an impairment in mitochondrial fatty acid β-oxidation. This includes long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency, first identified in 1989 (Wanders et al 1989) and now described in the literature in at least twelve additional patients. Identification of long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency in patients suspected to suffer from this enzyme defect is usually done in cultured skin fibroblasts

  • Long-chain 3-Hydroxyacyl-CoA Dehydrogenase in leukocytes and chorionic villus fibroblasts: potential for pre- and postnatal diagnosis.
    Journal of inherited metabolic disease, 1992
    Co-Authors: R. J. A. Wanders, L. Ijlst
    Abstract:

    In recent years an increasing number or inherited disorders or mitochondrial β-oxidation have been identified including long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency. So far, 11 patients have been described in the literature in which deficiency or long-chain 3-Hydroxyacyl-CoA Dehydrogenase has been proved enzymatically (Wanders et al 1989; Hale et al 1990; Rocchiccioli et al 1990; Wanders et al 1990; Carpenter et al 1991; Duran et al 1991; Jackson et al 1991; Dionisi-Vici et al 1991; Przyrembel et al 1991)

  • Long-chain 3-Hydroxyacyl-CoA Dehydrogenase deficiency.
    Journal of inherited metabolic disease, 1991
    Co-Authors: H. Przyrembel, C.a.j.m. Jakobs, L. Ijlst, J. B. C. De Klerk, R. J. A. Wanders
    Abstract:

    A new case of 3-Hydroxyacyl-CoA Dehydrogenase deficiency is described with a relatively benign course.

Alexander Steinbuchel - One of the best experts on this subject based on the ideXlab platform.

  • s 3 hydroxyacyl coa Dehydrogenase enoyl coa hydratase fadb from fatty acid degradation operon of ralstonia eutropha h16
    AMB Express, 2014
    Co-Authors: Elena Volodina, Alexander Steinbuchel
    Abstract:

    In this study (S)-3-Hydroxyacyl-CoA Dehydrogenase/enoyl-CoA hydratase (H16_A0461/FadB’, gene ID: 4247876) from one of two active fatty acid degradation operons of Ralstonia eutropha H16 has been heterologously expressed in Escherichia coli, purified as protein possessing a His-Tag and initially characterized. FadB’ is an enzyme with two catalytic domains exhibiting a single monomeric structure and possessing a molecular weight of 86 kDa. The C-terminal part of the enzyme harbors enoyl-CoA hydratase activity and is able to convert trans-crotonyl-CoA to 3-hydroxybutyryl-CoA. The N-terminal part of FadB’ comprises an NAD+ binding site and is responsible for 3-Hydroxyacyl-CoA Dehydrogenase activity converting (S)-3-hydroxybutyryl-CoA to acetoacetyl-CoA. Enoyl-CoA hydratase activity was detected spectrophotometrically with trans-crotonyl-CoA. (S)-3-Hydroxyacyl-CoA Dehydrogenase activity was measured in both directions with acetoacetyl-CoA and 3-hydroxybutyryl-CoA. FadB’ was found to be strictly stereospecific to (S)-3-hydroxybutyryl-CoA and to prefer NAD+. The K m value for acetoacetyl-CoA was 48 μM and V max 149 μmol mg−1 min−1. NADP(H) was utilized at a rate of less than 10% in comparison to activity with NAD(H). FadB’ exhibited optimal activity at pH 6–7 and the activity decreased at alkaline and acidic pH values. Acetyl-CoA, propionyl-CoA and CoA were found to have an inhibitory effect on FadB’. This study is a first report on biochemical properties of purified (S)-stereospecific 3-Hydroxyacyl-CoA Dehydrogenase/enoyl-CoA hydratase with the inverted domain order from R. eutropha H16. In addition to fundamental information about FadB’ and fatty acid metabolism, FadB’ might be also interesting for biotechnological applications.

  • (S)-3-Hydroxyacyl-CoA Dehydrogenase/enoyl-CoA hydratase (FadB’) from fatty acid degradation operon of Ralstonia eutropha H16
    AMB Express, 2014
    Co-Authors: Elena Volodina, Alexander Steinbuchel
    Abstract:

    In this study (S)-3-Hydroxyacyl-CoA Dehydrogenase/enoyl-CoA hydratase (H16_A0461/FadB’, gene ID: 4247876) from one of two active fatty acid degradation operons of Ralstonia eutropha H16 has been heterologously expressed in Escherichia coli, purified as protein possessing a His-Tag and initially characterized. FadB’ is an enzyme with two catalytic domains exhibiting a single monomeric structure and possessing a molecular weight of 86 kDa. The C-terminal part of the enzyme harbors enoyl-CoA hydratase activity and is able to convert trans-crotonyl-CoA to 3-hydroxybutyryl-CoA. The N-terminal part of FadB’ comprises an NAD+ binding site and is responsible for 3-Hydroxyacyl-CoA Dehydrogenase activity converting (S)-3-hydroxybutyryl-CoA to acetoacetyl-CoA. Enoyl-CoA hydratase activity was detected spectrophotometrically with trans-crotonyl-CoA. (S)-3-Hydroxyacyl-CoA Dehydrogenase activity was measured in both directions with acetoacetyl-CoA and 3-hydroxybutyryl-CoA. FadB’ was found to be strictly stereospecific to (S)-3-hydroxybutyryl-CoA and to prefer NAD+. The K m value for acetoacetyl-CoA was 48 μM and V max 149 μmol mg−1 min−1. NADP(H) was utilized at a rate of less than 10% in comparison to activity with NAD(H). FadB’ exhibited optimal activity at pH 6–7 and the activity decreased at alkaline and acidic pH values. Acetyl-CoA, propionyl-CoA and CoA were found to have an inhibitory effect on FadB’. This study is a first report on biochemical properties of purified (S)-stereospecific 3-Hydroxyacyl-CoA Dehydrogenase/enoyl-CoA hydratase with the inverted domain order from R. eutropha H16. In addition to fundamental information about FadB’ and fatty acid metabolism, FadB’ might be also interesting for biotechnological applications.