Tissue fractionation studies. 17. Intracellular distribution of monoamine oxidase, aspartate aminotransferase, alanine aminotransferase, <scp>d</scp>-amino acid oxidase and catalase in rat-liver tissue

Baudhuin, Pierre; Beaufay, Henri; Rahman-Li, Y.; Sellinger, O. Z.; Wattiaux, Robert; Jacques, P. J.; Duve, Christian de

doi:10.1042/bj0920179

Cited by 640 publications

(201 citation statements)

References 24 publications

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“…Adult flies of both sexes were quick-frozen on dry ice; abdominal organs were dissected either into 0.25 M sucrose/1% albumin to protect catalase activity for biochemical study (15) or into aldehyde fixative (see below) for microscopic study. Reproductive organs, eggs, and fat were removed as completely as possible and discarded.…”

Section: Methodsmentioning

confidence: 99%

“…Catalase activity was measured either in the whole homogenates or after the homogenates had been separated into subcellular fractions. These fractions were obtained by standard differential centrifugation methods using a Sorvall RC-5 centrifuge equipped with an SS-34 rotor (15,25). Fraction I was sedimented at 2700 x gamin; fraction II at 450,000 x gamin; fraction III was the supernatant from the second sedimentation step.…”

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confidence: 99%

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Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

Beard

Holtzman

1987

Proc. Natl. Acad. Sci. U.S.A.

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This study shows that peroxisomes are abundant in the Malpighian tubule and gut of wild-type Oregon R Drosophila melanogaster and that the peroxisomal population of the rosy-506 eye-color mutant differs from that of the wild type. Catalase activity in wild-type flies is demonstrable in bodies of appearance and centrifugal behavior comparable to the perixosomes of vertebrate tissues. Xanthine oxidase (xanthine:oxygen oxidoreductase, EC 1. Deficits in one or more of the peroxisomal enzymes and possible defects in peroxisomal structure are correlated with certain human inherited metabolic diseases (1-4). These diseases are still poorly understood, in part because different tissues show different effects of the mutation and in part because alterations in single genes can seemingly have multiple effects on the peroxisomes. Studies of the disorders are made difficult by their rareness and by the obvious limits of working with human material. It would be helpful to study analogues of the disorders in animals amenable to detailed analysis, but thus far such analogues have not been available. We chose to initiate our explorations for peroxisomal mutants in Drosophila by comparing wild-type Oregon R strain flies with the rosy-506 eye-color mutant, in which 90% of the structural gene for xanthine dehydrogenase (xanthine: NAD+ oxidoreductase, EC 1.1.1.204) is deleted (5, 6). This protein is a bifunctional oxidoreductase that can act as a dehydrogenase or as an oxidase (xanthine:oxygen oxidoreductase, EC 1.1.3.22) depending upon substrate and acceptor availability (7,8). The oxidase activity of the enzyme is critical to the degradation of purines in many organisms; the dehydrogenase function is central to the formation of pterinebased eye-color pigments in Drosophila. We suspected that this protein might be peroxisomal because (i) when acting as an oxidase, it produces hydrogen peroxide, a common feature of peroxisomal oxidases; (ii) it requires flavin in generating peroxide, as is found for many other peroxisomal oxidases; and (iii) its role in punine degradation is potentially functionally linked to activity of allantoicase and uricase, known peroxisomal enzymes (9). The subcellular localization of xanthine oxidase is not clearly established. Some cell fractionation studies show most of the enzyme to be soluble (9, 10), while other investigations report some xanthine oxidase cosedimenting with peroxisomal enzymes (11, 12).The study reported here identifies peroxisomes in the Malpighian tubule and gut of adult Drosophila wild-type Oregon R and rosy-506 strains, by virtue of their content of catalase, the peroxisomal "marker" enzyme. To localize xanthine oxidase, we have adapted cytochemical methods used to demonstrate other oxidases. With these methods, we have shown the presence of xanthine oxidase in peroxisomes of wild-type flies and the absence of this enzyme in rosy-506 flies. We also find signs that, as in the human mutations, a relatively simple genetic change can have complex effects on the peroxisomes. Prel...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

Beard

Holtzman

1987

Proc. Natl. Acad. Sci. U.S.A.

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“…These results can be compared with the observations of Baudhuin, Beaufay, Rahman-Li, Sellinger, Wattiaux, Jacques & de Duve (1964) on rat liver particles. These authors stated that, because of the small difference in sedimentation rate between mitochondria and lysosomes, a delicate procedure was required to obtain partial separation of these two types of particle.…”

Section: Discussionmentioning

confidence: 82%

The localization of lysosomal enzymes in chromaffin tissue

Smith¹,

Winkler²

1966

The Journal of Physiology

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SUMMARY1. An homogenate of bovine adrenal medulla contains significant amounts of six acid hydrolases: acid ribonuclease, acid deoxyribonuclease, cathepsin, acid phosphatase, /3-glucuronidase and arylsulphatase. Most of the activity of each enzyme could be sedimented in the large-granule fraction at 242,000 g-min.2. Differential centrifugation indicated the presence of three populations of particles, which sedimented at slightly different rates; these are, in order of decreasing sedimentation rate, mitochondria, particles containing the acid hydrolases, and chromaffin granules.3. The three types of particle could be separated by ultracentrifuging the large-granule fraction in a sucrose density gradient. Most of the activity of each hydrolase was recovered in a layer intermediate between those formed by mitochondria and chromaffin granules.4. The large-granule fraction therefore contains particles which are defined by their enzyme content as lysosomes.5. Highly purified chromaffin granules, containing less than 5 % of the activity of each acid hydrolase, were obtained from the gradient.

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“…(13): Maximum activity was obtained by preincubating the samples at 0°C for 2 h in a solution containing 1 mM sodium bicarbonate, 1 mM EDTA, 0 .001%' Triton-X-100, and 0.1% ethanol adjusted to pH 7 .4 with HCl . The assay mixture consisted of 18 mM imidazole-HC1 buffer, pH 7 .0, 1 mg/ml of bovine serum albumin (BSA), and 2 .67 mM H202 .…”

Section: Biochemical Analysesmentioning

confidence: 99%

Resolution of Three Distinct Populations of Nerve Endings From Rat Brain Homogenates by Zonal Isopycnic Centrifugation

Bretz¹,

Baggiolini²,

Häuser³

et al. 1974

The Journal of Cell Biology

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Conditions have been established for the fractionation of subcellular components of rat forebrain homogenates by zonal isopycnic equilibration in continuous sucrose density gradients using a B-XIV rotor . The fractions were analyzed biochemically and by ultrastructural morphometry . Starting from postnuclear supernates of forebrain homogenates, it has been possible to resolve three distinct populations of nerve endings from one another, as well as from free mitochondria and myelin fragments . The three types of nerve endings differ in their apparent specific gravity, their biochemical properties, and their ability selectively to accumulate exogenous transmitter substances in vitro . These three particle populations are likely to represent, in order of increasing modal equilibrium density, (a) cholinergic nerve endings, characterized by their high content of acetylcholine, (b) y-amino butyric acid (GABA) -containing nerve endings with high glutamate decarboxylase activity and the ability to accumulate exogenous GABA, (c) adrenergic nerve endings that accumulate exogenous dopamine and noradrenaline and exhibit high monoamine oxidase activity .

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Tissue fractionation studies. 17. Intracellular distribution of monoamine oxidase, aspartate aminotransferase, alanine aminotransferase, d-amino acid oxidase and catalase in rat-liver tissue

Cited by 640 publications

References 24 publications

Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

Peroxisomes in wild-type and rosy mutant Drosophila melanogaster.

The localization of lysosomal enzymes in chromaffin tissue

Resolution of Three Distinct Populations of Nerve Endings From Rat Brain Homogenates by Zonal Isopycnic Centrifugation

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