Adrenoleukodystrophy: Elevated C26 fatty acid in cultured skin fibroblasts

Moser, Hugo W.; Moser, Ann B.; Kawamura, Nariko; Murphy, Jerome V.; Suzuki, Kazuyuki; Schaumburg, Herbert H.; Kawaguchi, Yasuo

doi:10.1002/ana.410070607

Cited by 196 publications

(85 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The human cell lines were obtained and cultured as described by Moser et al (22). The transformed derivatives were kindly provided by Dr. S. J. Gould.…”

Section: Methodsmentioning

confidence: 99%

Cloning and Functional Expression of a Mammalian Gene for a Peroxisomal Sarcosine Oxidase

Reuber

Karl

Reimann

et al. 1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

Sarcosine oxidation in mammals occurs via a mitochondrial dehydrogenase closely linked to the electron transport chain. An additional H 2 O 2 -producing sarcosine oxidase has now been purified from rabbit kidney. A corresponding cDNA was cloned from rabbit liver and the gene designated sox. This rabbit sox gene encodes a protein of 390 amino acids and a molecular mass of 44 kDa identical to the molecular mass estimated for the purified enzyme. Sequence analysis revealed an N-terminal ADP-␤␣␤-binding fold, a motif highly conserved in tightly bound flavoproteins, and a C-terminal peroxisomal targeting signal 1. Sarcosine oxidase from rabbit liver exhibits high sequence homology (25-28% identity) to monomeric bacterial sarcosine oxidases. Both purified sarcosine oxidase and a recombinant fusion protein synthesized in Escherichia coli contain a covalently bound flavin, metabolize sarcosine, L-pipecolic acid, and L-proline, and cross-react with antibodies raised against L-pipecolic acid oxidase from monkey liver. Subcellular fractionation demonstrated that sarcosine oxidase is a peroxisomal enzyme in rabbit kidney. Transfection of human fibroblast cell lines and CV-1 cells (monkey kidney epithelial cells) with the sox cDNA resulted in a peroxisomal localization of sarcosine oxidase and revealed that the import into the peroxisomes is mediated by the peroxisomal targeting signal 1 pathway.In mammals a variety of H 2 O 2 -producing oxidases including D-amino-acid oxidase, D-aspartate oxidase, L-hydroxy-acid oxidase, acyl-CoA oxidase, and L-pipecolic acid oxidase are compartmentalized in peroxisomes. The H 2 O 2 generated from these reactions is then converted to H 2 O and O 2 by the peroxisomal matrix enzyme catalase (1). Several disorders have been described in which there is a defect in peroxisomal assembly that results in a partial or total absence of peroxisomal functions (for a review see Ref.2). Patients with these peroxisomal disorders such as Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum disease, and hyperpipecolatemia all have elevated levels of L-pipecolic acid, an imino acid, which in human and monkey liver is oxidized by a peroxisomal L-pipecolic acid oxidase (3). Indeed, L-pipecolic acid oxidase activity was not detected in liver samples from patients with Zellweger syndrome (4). Primates dehydrogenate L-pipecolic acid to ␦-piperideine-6-carboxylate which is spontaneously converted to ␣-aminoadipic acid ␥-semialdehyde.The subcellular localization of this pathway seems to differ in other mammalian species. In rabbits, guinea pigs, dogs, and sheep L-pipecolic acid oxidation is primarily mitochondrial (5). However, during our studies examining the subcellular distribution of L-pipecolic acid oxidation in rabbits, a considerable amount of L-pipecolic acid oxidation was detected in the peroxisomes, in addition to the previously reported mitochondrial activity (6). Interestingly, this peroxisomal enzyme showed a high specific activity for sarcosine and also oxidized L-pipecolic acid and L-p...

show abstract

“…The human cell lines were obtained and cultured as described by Moser et al (22). The transformed derivatives were kindly provided by Dr. S. J. Gould.…”

Section: Methodsmentioning

confidence: 99%

Cloning and Functional Expression of a Mammalian Gene for a Peroxisomal Sarcosine Oxidase

Reuber

Karl

Reimann

et al. 1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Concentrations of very long chain fatty acids in fibroblasts and plasma were measured by gas-liquid chromatography as previously described (26,27). The steps of plasmalogen biosynthesis in peroxisomes were assessed by the method of Roscher et al (28).…”

Section: Patient Selectionmentioning

confidence: 99%

Genetic and Phenotypic Heterogeneity in Disorders of Peroxisome Biogenesis—A Complementation Study Involving Cell Lines from 19 Patients

et al. 1989

View full text Add to dashboard Cite

“…A defect in adrenoleukodystrophy protein is known to be responsible for the X-linked neurodegeneration disorder, adrenoleukodystrophy. Adrenoleukodystrophy patient fibroblasts exhibit decreased ␤-oxidation of VLCFA, and VLCFA is accumulated in plasma and tissues (27,28). Furthermore, we have reported that PMP70 is involved in the metabolic transport of long-chain acyl-CoA across peroxisomal membranes, based on studies with CHO cells overexpressing wild-and mutant-type PMP70 (29).…”

mentioning

confidence: 99%

Identification of a Substrate-binding Site in a Peroxisomal β-Oxidation Enzyme by Photoaffinity Labeling with a Novel Palmitoyl Derivative

Kashiwayama

Tomohiro

Narita

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Peroxisomes play an essential role in a number of important metabolic pathways including ␤-oxidation of fatty acids and their derivatives. Therefore, peroxisomes possess various ␤-oxidation enzymes and specialized fatty acid transport systems. However, the molecular mechanisms of these proteins, especially in terms of substrate binding, are still unknown. In this study, to identify the substrate-binding sites of these proteins, we synthesized a photoreactive palmitic acid analogue bearing a diazirine moiety as a photophore, and performed photoaffinity labeling of purified rat liver peroxisomes. As a result, an 80-kDa peroxisomal protein was specifically labeled by the photoaffinity ligand, and the labeling efficiency competitively decreased in the presence of palmitoyl-CoA. Mass spectrometric analysis identified the 80-kDa protein as peroxisomal multifunctional enzyme type 2 (MFE2), one of the peroxisomal ␤-oxidation enzymes. Recombinant rat MFE2 was also labeled by the photoaffinity ligand, and mass spectrometric analysis revealed that a fragment of rat MFE2 (residues Trp 249 to Arg 251 ) was labeled by the ligand. MFE2 mutants bearing these residues, MFE2(W249A) and MFE2(R251A), exhibited decreased labeling efficiency. Furthermore, MFE2(W249G), which corresponds to one of the disease-causing mutations in human MFE2, also exhibited a decreased efficiency. Based on the crystal structure of rat MFE2, these residues are located on the top of a hydrophobic cavity leading to an active site of MFE2. These data suggest that MFE2 anchors its substrate around the region from Trp 249 to Arg 251 and positions the substrate along the hydrophobic cavity in the proper direction toward the catalytic center.Peroxisomes are organelles bound by a single membrane, which are present in almost all eukaryotic cells. Peroxisomes are involved in a variety of important metabolic processes including the ␤-oxidation of fatty acids, synthesis of plasmalogen, and bile acids (1). A specialized set of enzymes responsible for the peroxisomal functions are compartmentalized in the organelle, and the deficiency of peroxisomal enzymes causes severe metabolic disease such as acyl-CoA oxidase deficiency and D-bifunctional protein deficiency (2-4). Fibroblasts from patients with these deficiencies exhibit the disturbed peroxisomal ␤-oxidation of very long-chain fatty acids (VLCFAs), 4 and VLCFAs are accumulated in the plasma and tissues of these patients (5). Analyses of these peroxisomal diseases reflect the importance of the peroxisomal ␤-oxidation in organisms along with disclosing the enzymatic organization of the peroxisomal ␤-oxidation system. Peroxisomes are involved in the ␤-oxidation of VLCFAs, branched chain fatty acids such as pristanic acid, and bile acid intermediates such as dihydroxycholestanoic acid and trihydroxycholestanoic acid, which are incompatible with mitochondrial ␤-oxidation (6). Peroxisomal ␤-oxidation of fatty acids proceeds via a four-step pathway as in mitochondria, and multiple enzymes are involved in each step of the...

show abstract

Adrenoleukodystrophy: Elevated C26 fatty acid in cultured skin fibroblasts

Cited by 196 publications

References 24 publications

Cloning and Functional Expression of a Mammalian Gene for a Peroxisomal Sarcosine Oxidase

Cloning and Functional Expression of a Mammalian Gene for a Peroxisomal Sarcosine Oxidase

Genetic and Phenotypic Heterogeneity in Disorders of Peroxisome Biogenesis—A Complementation Study Involving Cell Lines from 19 Patients

Identification of a Substrate-binding Site in a Peroxisomal β-Oxidation Enzyme by Photoaffinity Labeling with a Novel Palmitoyl Derivative

Contact Info

Product

Resources

About