scholarly journals COMBINED BIOCHEMICAL AND MORPHOLOGICAL STUDY OF PARTICULATE FRACTIONS FROM RAT LIVER

1965 ◽  
Vol 26 (1) ◽  
pp. 219-243 ◽  
Author(s):  
Pierre Baudhuin ◽  
Henri Beaufay ◽  
Christian de Duve
Keyword(s):  
Electron Microscope ◽  
Amino Acid ◽  
Outer Membrane ◽  
Rat Liver ◽  
Morphological Study ◽  
Urate Oxidase ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Experimental Conditions ◽  
Dense Bodies

Six particulate preparations isolated from rat liver under different experimental conditions were analyzed biochemically and examined in the electron microscope. The results confirm the lysosomal nature of the pericanalicular dense bodies and demonstrate that the microbodies are the bearers of urate oxidase, catalase, and D-amino acid oxidase. Catalase, representing a major component of the particles, and D-amino acid oxidase appear to be associated with the structureless "sap" of the particles, urate oxidase with their crystalloid core or with their outer membrane.

scholarly journals THE SYNTHESIS AND TURNOVER OF RAT LIVER PEROXISOMES

1969 ◽  
Vol 41 (2) ◽  
pp. 521-535 ◽  
Author(s):  
Federico Leighton ◽  
Brian Poole ◽  
Paul B. Lazarow ◽  
Christian De Duve
Keyword(s):  
Amino Acid ◽  
Rat Liver ◽  
Isocitrate Dehydrogenase ◽  
Hydroxy Acid ◽  
Deae Cellulose ◽  
Urate Oxidase ◽  
Acid Oxidase ◽  
Total Proteins ◽  
Amino Acid Oxidase ◽  
Liver Peroxisomes

Rat liver peroxisomes isolated by density gradient centrifugation were disrupted at pH 9, and subdivided into a soluble fraction containing 90% of their total proteins and virtually all of their catalase, D-amino acid oxidase, L-α-hydroxy acid oxidase and isocitrate dehydrogenase activities, and a core fraction containing urate oxidase and 10% of the total proteins. The soluble proteins were chromatographed on Sephadex G-200, diethylaminoethyl (DEAE)-cellulose, hydroxylapatite, and sulfoethyl (SE)-Sephadex. None of these methods provided complete separation of the protein components, but these could be distributed into peaks in which the specific activities of different enzymes were substantially increased. Catalase, D-amino acid oxidase, and L-α-hydroxy acid oxidase contribute a maximum of 16, 2, and 4%, respectively, of the protein of the peroxisome. The contribution of isocitrate dehydrogenase could be as much as 25%, but is probably much less. After dissolution of the cores at pH 11 , no separation between their urate oxidase activity and their protein was achieved by Sephadex G-200 chromatography.

A quantitative histochemical study of D-amino acid oxidase activity in rat liver in relationship with feeding conditions.

1991 ◽  
Vol 39 (1) ◽  
pp. 81-86 ◽  
Author(s):  
H R Patel ◽  
W M Frederiks ◽  
F Marx ◽  
A J Best ◽  
C J Van Noorden
Keyword(s):  
Amino Acid ◽  
Rat Liver ◽  
Oxidase Activity ◽  
Specific Inhibitor ◽  
Physiological Role ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Specific Test ◽  
Final Reaction Product ◽  
Final Reaction

The histochemical method for the demonstration of D-amino acid oxidase activity in rat liver, based on the use of cerium ions and the diaminobenzidine-cobalt-hydrogen peroxide procedure, was improved by the application of unfixed cryostat sections and a semipermeable membrane interposed between section and gelled incubation medium. The amount of final reaction product precipitated in a granular form was about four times higher with this technique in comparison with conventional procedures using fixed sections and aqueous incubation media. The specificity of the reaction was proven by the 70% reduction of the amount of final reaction product when incubating in the presence of substrate and D,L-beta-hydroxybutyrate, a specific inhibitor of D-amino acid oxidase activity. Cytophotometric analysis of liver sections revealed that the specific test minus control reaction was linear with incubation time and section thickness. The Km value of the enzyme of 10.3 +/- 2.7 mM, as determined in periportal areas, is about five times the value found with biochemical methods in liver cell homogenates. The enzyme activity in periportal areas is about five times the activity in pericentral areas. Fasting (24 and 48 hr) induced a significant decrease in D-amino acid activity in periportal and pericentral areas. The possible physiological role of the enzyme in liver is discussed.

Heterogenous staining ofd-amino acid oxidase in peroxisomes of rat liver and kidney

1988 ◽  
Vol 88 (3-6) ◽  
pp. 277-285 ◽  
Author(s):  
S. Angermüller ◽  
H. D. Fahimi
Keyword(s):  
Amino Acid ◽  
Rat Liver ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Liver And Kidney

scholarly journals Immunoelectron microscopic study of a new D-amino acid oxidase-immunoreactive subcompartment in rat liver peroxisomes.

1991 ◽  
Vol 39 (1) ◽  
pp. 95-102 ◽  
Author(s):  
N Usuda ◽  
S Yokota ◽  
R Ichikawa ◽  
T Hashimoto ◽  
T Nagata
Keyword(s):  
Amino Acid ◽  
Rat Liver ◽  
Small Area ◽  
Electron Microscopic Observation ◽  
Glycolate Oxidase ◽  
Acid Oxidase ◽  
Electron Microscopic ◽  
Amino Acid Oxidase ◽  
The Matrix ◽  
Electron Lucent

We report the presence of a new subcompartment in rat liver peroxisomal matrix in which only D-amino acid oxidase is localized and other matrix enzymes are absent. By electron microscopic observation, the rat liver peroxisome has generally been considered to consist of a single limiting membrane, an electron-dense crystalline core, and a homogeneous matrix. Immunohistochemical staining for D-amino acid oxidase by the protein A-gold technique revealed the presence of a small area in the matrix that was immunoreactive for the enzyme and was less electron-dense than the surrounding matrix. The localization of D-amino acid oxidase in this small area of the peroxisomal matrix was confirmed by immunoelectron microscopy on freeze-substituted tissues processed without chemical fixation. To analyze the characteristics of the electron-lucent area, immunoreactivity for various peroxisomal enzymes, including catalase, acyl-CoA oxidase, enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase bifunctional protein, 3-ketoacyl-CoA thiolase, L-alpha-hydroxy acid oxidase (isozyme B), and glycolate oxidase (isozyme A), was assayed. The electron-lucent area was negative for all of these. By double staining for D-amino acid oxidase and catalase, using colloidal gold particles of different sizes, these enzymes were shown to be located in separate areas in the matrix.

scholarly journals Determination of D-Amino Acid Oxidase Activity in Tumour Cells

2002 ◽  
Vol 39 (6) ◽  
pp. 595-598 ◽  
Author(s):  
Taizo Sasamura ◽  
Akihiko Matsuda ◽  
Yukifumi Kokuba
Keyword(s):  
Amino Acid ◽  
Cancer Patients ◽  
Rat Liver ◽  
Rat Kidney ◽  
Tumour Cells ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Tissue Samples ◽  
Host Tissues

Background We evaluated the assay for determining D-amino acid oxidase (DAAO) activity in tumour cells, rat liver and rat kidney for studying the effects of D-amino acid-containing solution on cancer patients. Methods and Results In this method the amount of ammonia produced by the DAAO activity after removal of endogenous ammonia using a Sephadex G25 column was determined. The highest activity was observed in rat kidney, which was almost eight times that found in rat liver. As compared with host tissues, the DAAO activity in tumour cells was considerably less. Conclusions This DAAO assay may be useful for analysis of various tissue samples as well as tumour cells.

In situ kinetic measurements of d-amino acid oxidase in rat liver with respect to its substrate specificity

1993 ◽  
Vol 25 (8) ◽  
pp. 578-582 ◽  
Author(s):  
Wilma M. Frederiks ◽  
Cornelis J. F. Van Noorden ◽  
Frans Marx ◽  
Peter T. Gallagher ◽  
Brian P. Swann
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Rat Liver ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Kinetic Measurements

Oxidases and Oxygenases

2007 ◽  
Author(s):  
Perry A. Frey ◽  
Adrian D. Hegeman
Keyword(s):  
Amino Acid ◽  
Oxygen Atom ◽  
Phenylalanine Hydroxylase ◽  
Urate Oxidase ◽  
Galactose Oxidase ◽  
Ascorbate Oxidase ◽  
Acid Oxidase ◽  
Similar Reaction ◽  
Amino Acid Oxidase ◽  
Hydrolysis Of

An oxidase catalyzes the oxidation of a substrate by O2 without incorporating an oxygen atom into the product. A monooxygenase catalyzes oxidation by O2 with incorporation of one oxygen atom into the product, and oxidation by a dioxygenase proceeds with incorporation of both atoms of O2 into the product. These reactions generally require an organic or metallic coenzyme, with few exceptions, notably urate oxidase. Mechanisms of action of phenylalanine hydroxylase, galactose oxidase, and ascorbate oxidase are provided in chapter 4 in connection with the introduction of metallic coenzymes. In this chapter, we present cases of well-studied coenzyme and metal-dependent oxidases and oxygenases, and we consider one example of an oxidase that does not require a cofactor. Biochemical diversity may be a characteristic of oxidases, which include flavoproteins, heme proteins, copper proteins, and quinoproteins. The actions of copper and topaquinone-dependent amine oxidases are presented in chapter 3, and in chapter 4, two copper-dependent oxidases are discussed. In this chapter, we discuss flavin-dependent oxidases, a mononuclear iron oxidase, and a cofactor-independent oxidase. Flavin-dependent oxidases catalyze the reaction of O2 with an alcohol or amine to produce the corresponding carbonyl compound and H2O2. Examples include glucose oxidase, which produces gluconolactone and H2O2 from glucose and O2 according to. A D-Amino acid oxidase (EC 1.4.3.3) catalyzes a formally similar reaction to produce an α-keto acid from the corresponding α-D-amino acid. The oxidation of an amino acid by an oxidase produces ammonium ion in addition to hydrogen peroxide and the ketoacid, and so it is formally more complex. It proceeds in the three phases described in, the reduction of FAD to FADH2 by the amino acid, hydrolysis of the resultant α-iminoacid to the corresponding α-ketoacid and NH4, and oxidation of FADH2 by O2 to form H2O2. D-Amino acid oxidase is a thoroughly studied example of a flavoprotein oxidase. The enzyme is a 84-kDa homodimer containing one molecule of FAD per subunit. The mechanisms of the hydrolysis of imines and of the oxidation of dihydroflavins are discussed in chapters 1 and 3.

In situ kinetic measurements ofd-amino acid oxidase in rat liver with respect to its substrate specificity

1993 ◽  
Vol 25 (8) ◽  
pp. 578-582
Author(s):  
Wilma M. Frederiks ◽  
Cornelis J. F. Van Noorden ◽  
Frans Marx ◽  
Peter T. Gallagher ◽  
Brian P. Swann
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Rat Liver ◽  
Acid Oxidase ◽  
Amino Acid Oxidase ◽  
Kinetic Measurements
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