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R J Wanders

Publications and source records attributed to R J Wanders.

At least 199 records · Page 11Linked to original sources
PubMed1997

Cytosolic aspartate aminotransferase encoded by the AAT2 gene is targeted to the peroxisomes in oleate-grown Saccharomyces cerevisiae.

Fatty acid beta-oxidation in peroxisomes requires the continued uptake of fatty acids or their derivatives into peroxisomes and export of beta-oxidation products plus oxidation of NADH to NAD. In an earlier study we provided evidence for the existence of an NAD(H) redox shuttle in which peroxisomal malate dehydrogenase plays a pivotal role. In analogy to the NAD(H)-redox-shuttle systems in mitochondria we have investigated whether a malate/aspartate shuttle is operative in peroxisomes. The results described in this paper show that peroxisomes of oleate-grown Saccharomyces cerevisiae contain aspartate aminotransferase (AAT) activity. Whereas virtually all cellular AAT activity was peroxisomal in oleate-grown cells, we found that in glucose-grown cells most of the AAT activity resided in the cytosol. We demonstrate that the gene AAT2 codes for the cytosolic and peroxisomal AAT activities. Disruption of the AAT2 gene did not affect growth on oleate. Furthermore beta-oxidation of palmitate was normal. These results indicate that AAT2 is not essential for the peroxisomal NAD(H) redox shuttle.

Amino Acid Sequence↗
PubMed1997

Sterol carrier protein X (SCPx) is a peroxisomal branched-chain beta-ketothiolase specifically reacting with 3-oxo-pristanoyl-CoA: a new, unique role for SCPx in branched-chain fatty acid metabolism in peroxisomes.

One of the most important functions of peroxisomes, at least in humans, is the beta-oxidation of a range of different fatty acids and fatty acid derivatives. Recent studies have shown that the enzymatic machinery required for the beta-oxidations of these substrates, may be much more complex as originally thought. We now report that the conventional peroxisomal thiolase which has so far been thought to catalyze the thiolytic cleavage of the 3-oxoacyl-CoA esters of all fatty acids oxidized in peroxisomes, shows poor reactivity towards the 3-oxoacyl-CoA esters of 2-methyl branched-chain fatty acids such as pristanic acid. Our data further show, that SCPx, a 58 kDa protein with both thiolase and sterol carrier protein activity but unknown function so far, readily reacts with 3-oxopristanoyl-CoA. Taken together, our data show that SCPx plays a central role in branched chain fatty acid beta-oxidation in peroxisomes. This finding has major implications not only for the functional organization of the peroxisomal beta-oxidation system but also for studies dealing with the resolution of the underlying defect in patients with some defect in peroxisomal beta-oxidation.

Acetyl-CoA C-Acetyltransferase↗
PubMed1997

Complementation analysis of fibroblasts from peroxisomal fatty acid oxidation deficient patients shows high frequency of bifunctional enzyme deficiency plus intragenic complementation: unequivocal evidence for differential defects in the same enzyme protein.

In the last few years many patients have been reported with a defect in peroxisomal fatty acid beta-oxidation of unknown origin. Using a combined approach based on direct activity measurements of straight-chain acyl-CoA oxidase and complementation analysis after somatic cell fusion of fibroblasts, we have now classified 13 patients into 4 distinct groups representing different gene defects. Remarkably, we found intragenic complementation in group 2 so that group 2 is in fact made up of 3 distinct subgroups. The underlying basis for this peculiar phenomenon probably has to do with the fact that bifunctional protein harbors two catalytic activities including enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase. In group 2A enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase are defective whereas in group 2B and 2C either the hydratase or 3-hydroxyacyl-CoA dehydrogenase component of the bifunctional protein is deficient.

3-Hydroxyacyl CoA Dehydrogenases↗
PubMed1997

Succinyl-CoA:3-ketoacid coenzyme A transferase (SCOT): development of an antibody to human SCOT and diagnostic use in hereditary SCOT deficiency.

Succinyl-CoA:3-ketoacid CoA transferase (SCOT) is a key enzyme for ketone body utilization. Hereditary SCOT deficiency in humans (McKusick catalogue number 245050) is characterized by intermittent ketoacidotic attacks and permanent hyperketonemia. Since previously-available antibody to rat SCOT did not crossreact with human SCOT, we developed an antibody against recombinant human SCOT expressed in a bacterial system. The recombinant SCOT was insoluble except under denaturing conditions. Antibody raised to this polypeptide recognized denatured SCOT and proved useful for immunoblot analysis. On immunoblots, SCOT was easily detectable in control fibroblasts and lymphocytes but was detected neither in fibroblast extracts from four SCOT-deficient patients, nor in lymphocytes from two SCOT-deficient patients. These data indicate that immunoblot analysis is useful for diagnosis of SCOT deficiency in combination with enzyme assay.

Animals↗
PubMed1997

Studies on the intracellular localization of acetyl-CoA carboxylase.

The present work was performed to identify the subcellular localization of hepatic acetyl-CoA carboxylase (ACC). Cellular organelles involved in fatty acid oxidation that contain a malonyl-CoA sensitive carnitine palmitoyltransferase (CPT) activity or that are linked to the control of this activity were analysed for the presence of ACC. No significant amount of ACC was observed in the mitochondrial fraction prepared from isolated rat hepatocytes. Furthermore, no association of ACC activity and mass with isolated hepatic peroxisomes could be detected. Incubation of isolated hepatocytes with compounds known to affect the integrity of the cytoskeleton like okadaic acid or taxol indicates that ACC is associated with this subcellular structure of the hepatocyte. Such association may allow for efficient regulation of CPT activity and thus of fatty acid oxidation.

Acetyl-CoA Carboxylase↗
PubMed1997

New familial mitochondrial encephalopathy with macrocephaly, cardiomyopathy, and complex I deficiency.

Two siblings presented with a new phenotype consisting of fatal progressive macrocephaly and hypertrophic cardiomyopathy. Onset of symptoms started in both patients at the end of the first month of life with massive brain swelling causing macrocephaly and evolving to extensive brain destruction. Light microscopy of the lesions showed extensive small-vessel proliferation and gliosis. A distinct deficiency of complex I of mitochondrial respiratory chain was established in cultured fibroblasts, skeletal muscle, and heart muscle. Specific lack of complex I protein was demonstrated by two-dimensional gel electrophoresis.

Cardiomyopathies↗
PubMed1997

Succinyl-CoA:acetoacetate transferase deficiency: identification of a new patient with a neonatal onset and review of the literature.

UNLABELLED: We describe the clinical symptoms and biochemical findings of a patient with succinyl-CoA:acetoacetate transferase deficiency who presented in the neonatal period and review the current literature on this subject. Our patient was initially suspected to have distal renal tubular acidosis, and subsequently, a fasting test revealed severe metabolic ketoacidosis with normal blood glucose after 13 h which suggest a defect in ketolysis. In his cultured skin fibroblasts succinyl-CoA:acetoacetate transferase was deficient (residual activity 15%). Treatment in the acute phase consisted of sodium bicarbonate. At the present age of 9 years, psychomotor and physical development are within normal limits. CONCLUSION: Defects of ketolysis probably are underdiagnosed disorders and should be considered in infants and young children with persistent ketosis.

Acetoacetates↗
PubMed1997

A new peroxisomal beta-oxidation disorder in twin neonates: defective oxidation of both cerotic and pristanic acids.

Twin brothers were born with clinical symptoms indicating that they were suffering from Zellweger syndrome. However, instead of a generalized peroxisomal dysfunction, only very long-chain fatty acids and the pristanic acid/phytanic acid ratio were elevated in plasma and decreased oxidation of very long-chain fatty acids and pristanic acid was the only impairment found in fibroblasts. The other peroxisomal parameters tested were normal, including normal oxidation of phytanic acid and normal activity of dihydroxyacetonephosphate acyltransferase in fibroblasts as well as normal plasma bile acids. Although the biochemical results point to a defect in peroxisomal beta-oxidation, the isolated finding of impaired oxidation of very long-chain fatty acids and pristanic acid has to our knowledge not been reported previously and is difficult to explain by a deficiency of a known peroxisomal beta-oxidation enzyme.

Diseases in Twins↗
PubMed1997

Plasmalogens and oxidative stress: evidence against a major role of plasmalogens in protection against the superoxide anion radical.

Although ether-linked phospholipids have been known to be constituents of biological membranes for a long time, their physiological function has remained an enigma through the years. Inspired by the suggestion of Zoeller and coworkers that plasmalogens, which are specific types of ether-linked phospholipids characterized by the occurrence of an alpha, beta-unsaturated ether bond at the sn-1 position, are involved in the protection of cells against reactive oxygen species, we studied reactive oxygen species metabolism in cultured human skin fibroblasts. Menadione was used as intracellular generator of reactive oxygen species and cytochrome c as extracellular indicator for the production of reactive oxygen species. The finding that identical results were obtained in control and plasmalogen-deficient fibroblasts leads us to conclude that plasmalogens do not play a major role in protection against reactive oxygen species.

Anions↗