scholarly journals Cerebral-cortex hexokinase. Comparison of properties of solubilized mitochondrial and cytoplasmic activities

1970 ◽  
Vol 118 (1) ◽  
pp. 25-34 ◽  
Author(s):  
M. F. Thompson ◽  
H. S. Bachelard
Keyword(s):  
Cerebral Cortex ◽  
Deae Cellulose ◽  
Kinetic Properties ◽  
Sucrose Solutions ◽  
Michaelis Constants ◽  
Ammonium Sulphate Fractionation ◽  
Triton X ◽  
Two Peaks ◽  
Basic Similarity

1. Cerebral-cortex mitochondria, after purification by using high-density sucrose solutions, were extracted with Triton X-100. The total hexokinase activity of the intact mitochondria was increased by 50–80% in the Triton extracts. 2. Triton X-100 was removed from mitochondrial extracts by a combination of ammonium sulphate fractionation and DEAE-cellulose chromatography. Mitochondrial hexokinase remained soluble after removal of extractant. 3. The behaviour of solubilized mitochondrial hexokinase was compared with soluble cytoplasmic hexokinase from the same samples of cerebral cortex on identical columns of DEAE-cellulose. Two peaks were eluted from each source of hexokinase. The distribution between hexokinase peaks was similar for the two sources. Peak I (approx. 80% of the total hexokinase) from each was eluted at identical concentrations of potassium chloride and slight differences were observed in the elution profiles for peak II. 4. The purified mitochondrial hexokinase showed the following kinetic properties: peak I, Km(ATP) 0.60mm, Km(glucose) 0.042mm; peak II, Km(ATP) 0.66mm, Km(glucose) 0.043mm. The purified cytoplasmic hexokinase Michaelis constants were: peak I, Km(ATP) 0.56mm, Km(glucose) 0.048mm; peak II, Km(ATP) 0.68mm, Km(glucose) 0.062mm. 5. Although no significant differences between mitochondrial and cytoplasmic hexokinases were noted in chromatographic behaviour or in the kinetic properties studied, the purified mitochondrial enzyme was activated slightly (approx. 20%) by Triton X-100, in contrast with the cytoplasmic enzyme, which was not affected. 6. The results, taken to indicate basic similarity between mitochondrial and cytoplasmic hexokinases, are discussed in relation to the role of the two sources of enzyme in the metabolism of the tissue.

scholarly journals The characterization of two reduced nicotinamide–adenine dinucleotide phosphate-linked aldehyde reductases from pig brain

1972 ◽  
Vol 130 (3) ◽  
pp. 765-772 ◽  
Author(s):  
A. J. Turner ◽  
K. F. Tipton
Keyword(s):  
Deae Cellulose ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Aldehyde Reductase ◽  
Enzyme Preparations ◽  
Pig Brain ◽  
Michaelis Constants ◽  
Probable Role ◽  
Reduced Nicotinamide Adenine Dinucleotide

1. NADPH-linked aldehyde reductase from pig, ox and rat brain exhibits non-linear reciprocal plots when partially purified enzyme preparations are studied. 2. In pig brain this non-linearity is due to the presence of two distinct aldehyde reductases, which can be separated by DEAE-cellulose chromatography. 3. These two enzymes can be distinguished by several criteria, including pH optima, Michaelis constants for substrates and their inhibitor sensitivity. 4. The probable role of these enzymes in the metabolism of the aldehydes derived from the biogenic amines is discussed.

Partial purification and characterization of the lipase of a facultatively psychrophilic bacterium (Acinetobacter O16)

1975 ◽  
Vol 21 (4) ◽  
pp. 434-441 ◽  
Author(s):  
Colette Breuil ◽  
D. J. Kushner
Keyword(s):  
Deae Cellulose ◽  
Crude Enzyme ◽  
Partial Purification ◽  
Psychrophilic Bacterium ◽  
Sephadex Column ◽  
Triton X ◽  
Two Peaks ◽  
Temperature Optima ◽  
Cellulose Column

The extracellular lipase(s) of the psychrophile Acinetobacter O16 was studied. When the enzyme was precipitated by (NH4)2SO4 and passed through a Sephadex G 200 column, two peaks of lipase activity appeared. The larger peak, which behaved like a substance of high molecular weight, being eluted in the void volume, was purified 250-fold over the crude enzyme (culture supernatant) by passage through a DEAE Sephadex column. When the enzyme was applied to a DEAE-cellulose column it could not be eluted unless it had first been treated with the detergent Triton X 100. It is suggested that lipids or phospholipids make up an important part of the molecule.The activity of the crude and partly purified enzymes was studied in relation to pH and temperature optima. Lipases from the psychrophilic Acinetobacter O16 and from the mesophilic Acinetobacter O4 reacted in the same way to temperature. The crude enzyme from Acinetobacter O16 was more temperature-stable than the purified enzyme.

scholarly journals The separation, partial purification and some properties of isoenzymes of aldolase from guinea-pig cerebral cortex

1969 ◽  
Vol 112 (5) ◽  
pp. 587-594 ◽  
Author(s):  
P. C. Nicholas ◽  
H S Bachelard
Keyword(s):  
Cerebral Cortex ◽  
Guinea Pig ◽  
Deae Cellulose ◽  
Partial Purification ◽  
Subunit Structure ◽  
Starch Gel Electrophoresis ◽  
Activity Ratios ◽  
Lysine Residues ◽  
Starch Gel ◽  
Ammonium Sulphate Fractionation

1. Aldolase isoenzymes from guinea-pig cerebral cortex were partially purified and separated by ammonium sulphate fractionation and chromatography on DEAE-cellulose. 2. Each purified isoenzyme was shown to be virtually uncontaminated with other forms by starch-gel electrophoresis. The quantitative distribution of the isoenzymes was: I, 6·2%; II, 5·2%; III, 15·3%; IV, 25·7%; V, 33·3%. 3. The pH optima for the five separated isoenzymes were similar; all were in the range pH7·5–8·0. Values for pKa (6·31–6·55) and pKb (9·45–9·59) were calculated from the data and suggested the involvement of histidine and lysine residues. 4. The stabilities of the isoenzymes were shown to be I=II>III>IV>V at pH4·4 in order of decreasing stability and are discussed in terms of subunit structure. 5. The substrate activity ratios (fructose 1,6-diphosphate/fructose 1-phosphate) were measured and all were in the range 12–44.

scholarly journals Isolation, characterization and the role of rabbit testicular arysulphatase A in fertilization

1979 ◽  
Vol 181 (2) ◽  
pp. 331-337 ◽  
Author(s):  
A A Farooqui ◽  
P N Srivastava
Keyword(s):  
Specific Activity ◽  
Deae Cellulose ◽  
Cumulus Cells ◽  
Kinetic Properties ◽  
Neutral Sugar ◽  
Acetylneuraminic Acid ◽  
Crude Homogenate ◽  
Step Procedure ◽  
Arylsulphatase A

Arysulphatase A was purified from rabbit testis. The purification was accomplished by a four-step procedure involving (NH4)2SO4 fractionation, chromatography on DEAE-cellulose, SP(sulphopropyl)-Sephadex and affinity chromatography on concanavalin A-Sepharose. The specific activity of purified preparation was 135 mumol/min per mg of protein, which represented an increase of 900-fold above that of the crude homogenate. The purified enzyme (20-50 micrograms) was found to move electrophoretically as a single band on polyacrylamide gel at pH 7.2 and 8.4. The homogeneous enzyme was shown to be a glycoprotein with 0.8% (w/w) of N-acetylneuraminic acid and 20% neutral sugar. The treatment of purified enzyme with bacterial neuraminidase had no effect on enzyme activity or kinetic properties, but it changed the elution prolife of rabbit testis arylsulphatase A through DEAE-Sephadex. The purified enzyme was strongly inhibited by Cu2+, Fe3+ and Ag+. It hydrolysed several sulphate esters including cerebroside 3-sulphate, ascorbic acid 2-sulphate and steroid sulphates. Pure arysulphatase was effective in dispersing the cumulus cells of rabbit ova.

The Activation of Human Factor X in Sodium Citrate: the Role of Factor VII

1976 ◽  
Vol 36 (01) ◽  
pp. 104-114 ◽  
Author(s):  
D. L Aronson ◽  
A. J Mustafa
Keyword(s):  
Sodium Citrate ◽  
Human Factor ◽  
Deae Cellulose ◽  
Factor Ix ◽  
Factor Vii ◽  
Factor X

SummaryHuman factor X was purified by several different procedures yielding products which had varying amounts of factor VII and factor IX. Treatment with CHC13 during the fractionation of the factor X removed 95% of the factor VII and factor IX activity and the resulting factor X activated more slowly when incubated in 25% sodium citrate. Removal of residual factor VII by DEAE cellulose chromatography yielded a factor X which activated still more slowly and less completely. When the factor VII, removed by chromatography, was added to the chromatographed factor X, the ability to be activated in 25% sodium citrate was restored. Confirmatory evidence for the role of factor VII in this reaction was the inhibition of the conversion of the factor X by both DFP and SBTI.

Chromatographic Isolation of the LDH-Isoenzymes of Human Blood Platelets and an Investigation of Their Enzyme Kinetics

1968 ◽  
Vol 20 (03/04) ◽  
pp. 301-313 ◽  
Author(s):  
W Schneider ◽  
K Schumacher ◽  
B Thiede ◽  
R Gross
Keyword(s):  
Human Blood ◽  
Blood Platelets ◽  
Deae Cellulose ◽  
Order Reaction ◽  
Kinetic Properties ◽  
Ldh Isoenzymes ◽  
Kinetic Investigations ◽  
Substrate Concentrations ◽  
Chromatographic Isolation ◽  
Human Blood Platelets

SummaryThe LDH-isoenzymes of human blood platelets show a distinct predominance of the isoenzymes 2 and 3 upon chromatography on DEAE-cellulose. Small amounts of LDH-1 are also present, while only traces of LDH-4 and -5 can be detected.Enzyme kinetic investigations of the principal isoenzymes LDH-1, -2 and -3 clearly show that the differences in inhibition constants with pyruvate as substrate which are demonstrable at 25° largely disappear at 37°. On the other hand, the differences among the isoenzymes in their affinity for pyruvate and lactate as substrate as well as in with respect to the optimal substrate concentrations of pyruvate are more marked at 37° than at 25°. Also, the type of inhibition found with lactate as substrate is increasingly the expression of a higher order reaction in going from LDH-1 to LDH-3. A dependence of the LDH distribution pattern upon the metabolism of the cell is discussed. A comparison of our results with thrombocytes with those of other workers with erythrocytes and leucocytes makes it unlikely that the LDH pattern is directly dependent upon the existence of an oxidative metabolism. Rather, the redox potential of the cell could be of importance for the nature of the pattern of isoenzymes and for their differing kinetic properties.

scholarly journals Membrane-surfactant interactions. The role of surfactant in mitochondrial complex III-phospholipid-Triton X-100 mixed micelles.

1986 ◽  
Vol 261 (14) ◽  
pp. 6578-6584
Author(s):  
J M Valpuesta ◽  
J L Arrondo ◽  
M C Barbero ◽  
M Pons ◽  
F M Goñi
Keyword(s):  
Mixed Micelles ◽  
Complex Iii ◽  
Mitochondrial Complex ◽  
Surfactant Interactions ◽  
Triton X

scholarly journals Pre-motor versus motor cerebral cortex neuromodulation for chronic neuropathic pain

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Igor Lavrov ◽  
Timur Latypov ◽  
Elvira Mukhametova ◽  
Brian Lundstrom ◽  
Paola Sandroni ◽  
...  
Keyword(s):  
Neuropathic Pain ◽  
Cerebral Cortex ◽  
Motor Cortex ◽  
Initial Assessment ◽  
Motor Cortex Stimulation ◽  
Chronic Neuropathic Pain ◽  
Long Term Effect ◽  
Stimulation Of

AbstractElectrical stimulation of the cerebral cortex (ESCC) has been used to treat intractable neuropathic pain for nearly two decades, however, no standardized approach for this technique has been developed. In order to optimize targeting and validate the effect of ESCC before placing the permanent grid, we introduced initial assessment with trial stimulation, using a temporary grid of subdural electrodes. In this retrospective study we evaluate the role of electrode location on cerebral cortex in control of neuropathic pain and the role of trial stimulation in target-optimization for ESCC. Location of the temporary grid electrodes and location of permanent electrodes were evaluated in correlation with the long-term efficacy of ESCC. The results of this study demonstrate that the long-term effect of subdural pre-motor cortex stimulation is at least the same or higher compare to effect of subdural motor or combined pre-motor and motor cortex stimulation. These results also demonstrate that the initial trial stimulation helps to optimize permanent electrode positions in relation to the optimal functional target that is critical in cases when brain shift is expected. Proposed methodology and novel results open a new direction for development of neuromodulation techniques to control chronic neuropathic pain.

scholarly journals The Possible Role of Radiative Acceleration in Supporting Extended Atmospheres in Be Stars

1976 ◽  
Vol 70 ◽  
pp. 377-382 ◽  
Author(s):  
R. L. Kurucz ◽  
R. E. Schild
Keyword(s):  
Effective Temperature ◽  
Detailed Calculation ◽  
Be Stars ◽  
Be Star ◽  
Hα Emission ◽  
Two Peaks ◽  
Radiative Acceleration ◽  
Emission Strength ◽  
Peaked Function

A detailed calculation of the radiative acceleration in B-type stars shows it to be a double-peaked function of effective temperature at small optical depths. The two peaks are shown to coincide approximately with peaks in the distribution of mean Hα emission strength as a function of B - V color in Be stars. These facts suggest that radiation may play an important role in the support of the Be star extended atmosphere.

Role of Cerebral Cortex in Voluntary Movements

1985 ◽  
Vol 65 (5) ◽  
pp. 624-635 ◽  
Author(s):  
Paul D. Cheney
Keyword(s):  
Cerebral Cortex ◽  
Voluntary Movements
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