CYTOCHROME OXIDASE ACTIVITY OF CELL NUCLEI

1954 ◽  
Vol 32 (1) ◽  
pp. 548-552 ◽  
Author(s):  
David Rubinstein ◽  
Orville F. Denstedt
Keyword(s):  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Oxidase Activity ◽  
Nuclear Material ◽  
Liver Cells ◽  
Nuclear Fraction ◽  
Cell Nuclei ◽  
Nuclear Membranes ◽  
Rat Liver Cells ◽  
Chicken Erythrocytes

The presence of cytochrome oxidase in the nuclear material from chicken erythrocytes and rat-liver cells has been demonstrated with the aid of p-phenylenediamine as the agent for reducing cytochrome c. The three reagents p-phenylenediamine, ascorbate, and hydroquinone are effective in the estimation of cytochrome oxidase in mitochondrial preparations, but only the first mentioned agent can effect the reduction of endogenous cytochrome c. The addition of cytochrome c will increase the oxidase activity of the liver-cell mitochondria but not of the nuclear fraction from the erythrocytes or the liver cells, presumably because of the impermeability of the nuclear membranes to added cytochrome c.

CYTOCHROME OXIDASE ACTIVITY OF CELL NUCLEI

1954 ◽  
Vol 32 (5) ◽  
pp. 548-552 ◽  
Author(s):  
David Rubinstein ◽  
Orville F. Denstedt
Keyword(s):  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Oxidase Activity ◽  
Nuclear Material ◽  
Liver Cells ◽  
Nuclear Fraction ◽  
Cell Nuclei ◽  
Nuclear Membranes ◽  
Rat Liver Cells ◽  
Chicken Erythrocytes

The presence of cytochrome oxidase in the nuclear material from chicken erythrocytes and rat-liver cells has been demonstrated with the aid of p-phenylenediamine as the agent for reducing cytochrome c. The three reagents p-phenylenediamine, ascorbate, and hydroquinone are effective in the estimation of cytochrome oxidase in mitochondrial preparations, but only the first mentioned agent can effect the reduction of endogenous cytochrome c. The addition of cytochrome c will increase the oxidase activity of the liver-cell mitochondria but not of the nuclear fraction from the erythrocytes or the liver cells, presumably because of the impermeability of the nuclear membranes to added cytochrome c.

Reduction of Cytochrome c by Tetrathionate in the Presence of Tetrathionate Hydrolase Purified from Sulfur-Grown Acidithiobacillus Ferrooxidans ATCC 23270

2009 ◽  
Vol 71-73 ◽  
pp. 243-246
Author(s):  
Taher M. Taha ◽  
Fumiaki Takeuchi ◽  
Tsuyoshi Sugio
Keyword(s):  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Acidithiobacillus Ferrooxidans ◽  
Oxidase Activity ◽  
Elemental Sulfur ◽  
Enzyme System ◽  
Size Exclusion ◽  
Ammonium Sulfate Precipitation ◽  
Interesting Phenomenon ◽  
Oxidase Enzyme

It is mysterious that, when A. ferrooxidans ATCC 23270 cells grow on elemental sulfur, they have high iron oxidase activity comparable to that of iron-grown cells as well as high activities of sulfide:ferric ion oxidoreductase (SFORase) and tetrathionate hydrolase. To clarify this interesting phenomenon, cytochrome c and tetrathionate hydrolase were purified from sulfur-grown A. ferrooxidans cells using ammonium sulfate precipitation, Phenyl column chromatography, and SuperdexTM 75 and Sephadex G-100 size exclusion column chromatographies. The purified cytochrome c was reduced by tetrathionate in the presence of purified tetrathionate hydrolase, but not in the absence of the enzyme. When the partially purified cytochrome c fraction containing aa3-type cytochrome oxidase was used, both cytochrome c and aa3-type cytochrome oxidase were reduced by tetrathionate in the presence of purified tetrathionate hydrolase. These results indicate that tetrathionate in the presence of tetrathionate hydrolase can reduce iron oxidase enzyme system containing cytochrome c and aa3-type cytochrome oxidase as tetrathionate hydrolase decomposes tetrathionate to produce thiosulfate, elemental sulfur, and sulfate; and the formed thiosulfate can chemically reduce cytochrome c and Fe3+.

Lectin receptor sites on rat liver cell nuclear membranes

1976 ◽  
Vol 22 (2) ◽  
pp. 335-344
Author(s):  
I. Virtanen ◽  
J. Wartiovaara
Keyword(s):  
Rat Liver ◽  
Liver Cell ◽  
Liver Cells ◽  
Nuclear Surface ◽  
Con A ◽  
Lectin Receptor ◽  
Nuclear Membranes ◽  
Cytoplasmic Surface ◽  
Receptor Sites ◽  
Rat Liver Cells

The presence and localization of lectin receptor sites on rat liver cell nuclear and other endomembranes was studied by light and electron microscopy using fluorescein and ferritin-coupled lectin conjugates. Isolated nuclei labelled with fluorescein-conjugated Concanavalin A (Con A) or wheat germ agglutinin (WGA) often showed membrane staining, which sometimes was especially bright on small stretches of the nuclear surface. Unlabelled nuclei and nuclei with a complete ring fluorescence were also seen. The nuclear fluorescence corresponded in intensity to that seen on the surface of isolated rat liver cells. Con A-ferritin particles were seldom detected on the cytoplasmic surface of the intact nuclear envelope. However, at places where the 2 leaflets of the envelope were widely separated or where the outer nuclear membrane was partly torn away, heavy labelling was seen on the cisternal surface of both the inner and outer nuclear membranes. Labelling with Con A-ferritin was also found on the cisternal side of rough endoplasmic reticulum present in the specimens. No labelling was seen on the cytoplasmic surface of mitochondrial outer membrane. The results demonstrate the presence of binding sites for Con A and WGA in nuclei and an asymmetric localization of these sites on the cisternal side of ribosome-carrying endomembranes in rat liver cells.

scholarly journals Localization and biosynthesis of NADH-cytochrome b5 reductase, an integral membrane protein, in rat liver cells. I. Distribution of the enzyme activity in microsomes, mitochondria, and golgi complex.

1980 ◽  
Vol 85 (3) ◽  
pp. 501-515 ◽  
Author(s):  
N Borgese ◽  
J Meldolesi
Keyword(s):  
Cytochrome C ◽  
Rat Liver ◽  
Golgi Complex ◽  
Cytochrome B5 ◽  
Liver Cells ◽  
Cytochrome C Reductase ◽  
Cytochrome B5 Reductase ◽  
Galactosyl Transferase ◽  
Rat Liver Cells ◽  
Nadh Cytochrome

The subcellular distribution of NADH-cytochrome b5 reductase in rat liver cells was reinvestigated. In fresh heavy and light Golgi fractions (GF3 and GF1 + 2) and in mitochondria, the specific activity of rotenone-insensitive NADH-cytochrome c reductase was approximately 100, 60, and 30%, respectively, of the value found in microsomes. However, the Golgi enzyme was unstable inasmuch as pelleting and resuspending the fresh fractions resulted in a considerable inactivation (40--60%), which was further increased with subsequent storage at 4 degrees C. A similar inactivation was observed using cytochrome b5 but not ferricyanide as electron acceptor. The inactivation of Golgi NADH-cytochrome c reductase activity was independent of the protein concentration of the fractions during storage, was unaffected by the addition of the antioxidant butylated hydroxytoluene, but was partly prevented by buffering the fractions at neutral pH and by storage at--20 degrees C. A total Golgi fraction was analyzed by density equilibration on continuous sucrose gradients after exposure to digitonin. As expected, the distribution of both protein and galactosyl transferase were shifted to higher densities by this treatment. However, not all galactosyl transferase-bearing elements were shifted to the same extent by exposure to the detergent, suggesting a biochemical heterogeneity of the Golgi complex. In contrast to their behavior in microsomes, the distribution of NADH-cytochrome c reductase and cytochrome b5 of Golgi fractions was shifted by digitonin, although to a lesser extent than that of galactosyl transferase. These results indicate that NADH-cytochrome b5 reductase is an authentic component of Golgi membranes, as well as of microsomes and of mitochondria. The conflicting results reported in the past on the Golgi localization of the enzyme could be due, on the one hand, to the differential lability of the activity in its various subcellular locations and, on the other, to the heterogeneity of the Golgi complex in terms of both cholesterol and enzyme distribution.

scholarly journals Modulation of cytochrome oxidase activity by inorganic and organic phosphate

1987 ◽  
Vol 248 (1) ◽  
pp. 161-165 ◽  
Author(s):  
F Malatesta ◽  
G Antonini ◽  
P Sarti ◽  
M Brunori
Keyword(s):  
Ionic Strength ◽  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Oxidase Activity ◽  
Respiratory Control ◽  
Organic Phosphate ◽  
Phospholipid Vesicles ◽  
Respiratory Control Ratio ◽  
Phospholipid Membrane ◽  
Ferrocytochrome C

The activity of cytochrome oxidase reconstituted into phospholipid vesicles has been studied as a function of orthophosphate, ATP and inositol hexakisphosphate concentrations. The respiratory-control ratio was found to be quite sensitive to these compounds and was inversely related to the anion concentration. This effect is related to a phosphate-dependent decrease in the rate constant for ferrocytochrome c oxidation observed in the presence of ionophores. The data cannot be interpreted simply on the basis of ionic strength, which is known to limit cytochrome c binding to cytochrome oxidase, since cytochrome oxidase-containing vesicles responded differently to phosphate depending on the energization state of the phospholipid membrane.

scholarly journals Equilibrium relations between the oxidation—reduction reactions and the adenosine triphosphate synthesis in suspensions of isolated liver cells

1974 ◽  
Vol 140 (1) ◽  
pp. 57-64 ◽  
Author(s):  
David F. Wilson ◽  
Marion Stubbs ◽  
Richard L. Veech ◽  
Maria Erecińska ◽  
Hans A. Krebs
Keyword(s):  
Cytochrome C ◽  
Adenine Nucleotides ◽  
Atp Synthesis ◽  
Mitochondrial Respiratory Chain ◽  
Liver Cells ◽  
Inner Mitochondrial Membrane ◽  
Phosphorylation State ◽  
Oxidation Reduction ◽  
Rat Liver Cells ◽  
The Individual

1. The redox state of cytochrome c, cytochrome a and the mitochondrial NAD couple, and the phosphorylation state of the adenine nucleotides, were measured in suspensions of isolated rat liver cells. 2. The ΔG for the transfer of two electrons from the mitochondrial NAD to the cytochrome c couple is calculated to be 104kJ (24.8kcal). 3. The ΔG associated with the synthesis of ATP at the measured phosphorylation state is calculated to be 95kJ (22.7kcal)/2mol of ATP. 4. The near equality of ΔG of the electron-transport process and ΔG required for ATP synthesis indicates near-equilibrium between the mitochondrial respiratory chain and the extramitochondrial phosphorylation state. 5. The existence of near-equilibrium in the coupled reactions implies that the respiratory activity depends on the ratio [ATP]/[ADP][Pi] and not on the concentrations of the individual reactants. 6. If the overall system of oxidative phosphorylation is at near-equilibrium, all intermediary reactions must also be at equilibrium. Hence if the intramitochondrial and extramitochondrial phosphorylation states are indeed different, it follows that any differences in the activities of ATP, ADP and Pi must be coupled to ion gradients and/or potentials across the inner mitochondrial membrane in such a way that translocation occurs without loss of free energy. 7. The metabolic state of the mitochondria in the cell can be defined by the turnover number of the cytochromes, the cytoplasmic phosphorylation state, and the oxidation–reduction potential of the NAD couple, rather than by the availability of ADP, substrate and O2.

scholarly journals RESPIRATORY ENZYMES AND MITOCHONDRIAL MORPHOLOGY OF HELA AND L CELLS TREATED WITH CHLORAMPHENICOL AND ETHIDIUM BROMIDE

1972 ◽  
Vol 53 (1) ◽  
pp. 127-142 ◽  
Author(s):  
Mary E. King ◽  
Gabriel C. Godman ◽  
Donald W. King
Keyword(s):  
Cytochrome C ◽  
Cytochrome Oxidase ◽  
Ethidium Bromide ◽  
Enzyme Activities ◽  
Oxidase Activity ◽  
Mitochondrial Morphology ◽  
Respiratory Enzymes ◽  
Cytochrome Oxidase Activity ◽  
Control Value ◽  
L Cells

Exposure of HeLa and L cells to chloramphenicol causes a progressive dose-dependent decrease in cytochrome oxidase and succinate-cytochrome c reductase activities, concomitant with an increase in the amount of cytochrome c. At 2–3 days, the specific activities of the enzymes have fallen to about one-half of control values; the mitochondria appear swollen. By day 5, enzyme activities are about one-quarter of control values; the mitochondria are more swollen, with disorientation and disintegration of cristae. By day 6–8, after three generations, growth has stopped, enzyme activities are approximately the same as on day 5, and cytochrome c content has reached 170% of control value. Mitochondria show severe changes, cristae being affected more than peripheral inner membrane. The number of profiles continues to be nearly normal. After 30 days, cytochrome oxidase activity remains low but now there are mitochondria in intermediate and condensed configuration. There is a gradual accumulation in the cytoplasm of smooth membrane elements. If chloramphenicol is removed, cells recover. Ethidium bromide treatment for up to 8 days yields results virtually identical to those obtained with chloramphenicol. Cells treated with 10-4 M KCN show a decrease in cytochrome oxidase activity to about one-third of control value and an elevated amount of cytochrome c. Only a small number of mitochondria appear damaged. Autochthonous mitochondrial syntheses appear to be essential for the organization of the cristae. When cytochrome oxidase activity is impaired, a regulatory mechanism for cytochrome biosynthesis geared to mitochondrial function may be lacking, resulting in an increase in cytochrome c content.

scholarly journals A reliable Feulgen-acriflavine-SO2 staining procedure for quantitative DNA measurements.

1980 ◽  
Vol 28 (9) ◽  
pp. 1007-1013 ◽  
Author(s):  
H J Tanke ◽  
E M van Ingen
Keyword(s):  
Rat Liver ◽  
Liver Cells ◽  
Staining Procedure ◽  
Rat Liver Cells ◽  
Scanning Cytophotometry ◽  
Chicken Erythrocytes ◽  
Dna Measurements

Feulgen-acriflavine-SO2 staining was performed on chicken erythrocytes and rat liver cells under different cytochemical conditions. The obtained results were compared to conventional Feulgen-pararosaniline-SO2 staining by means of scanning cytophotometry and microfluorometry. It was found that if acid ethanol rinsing was included in the Feulgen-acriflavine-SO2 procedure, both Feulgen procedures were fully comparable in specificity and quantitative aspects.

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