scholarly journals Metabolism of the herbicide 2,6-dichlorobenzonitrile in rabbits and rats

1966 ◽  
Vol 101 (3) ◽  
pp. 698-706 ◽  
Author(s):  
JG Wit ◽  
H Van Genderen
Keyword(s):  
Oral Administration ◽  
Urinary Excretion ◽  
Rat Liver ◽  
High Speed ◽  
Liver Homogenate ◽  
Acid Formation ◽  
Thiol Compounds ◽  
Enzyme Preparations ◽  
Phenolic Metabolites ◽  
High Speed Supernatant

1. The metabolism of 2,6-dichlorobenzonitrile was studied in rabbits and rats. Oral administration caused an increased urinary excretion of glucuronides and ethereal sulphates. There was also an indication of mercapturic acid formation. 2,6-Dichloro-3-hydroxybenzonitrile and its 4-hydroxy analogue were identified as metabolites in the urine. A small amount of the unchanged substance was recovered from the faeces. 2. By using 2,6-dichlorobenzo[(14)C]nitrile the phenolic metabolites were determined quantitatively and some other possible metabolic routes were excluded. 3. Incubation of 2,6-dichlorobenzonitrile with enzyme preparations (papain and high-speed supernatant of rat-liver homogenate plus glutathione) gave no indications for a reaction with thiol compounds.

‘Binding’ of [14C]phenol to rat liver high-speed supernatant

1980 ◽  
Vol 8 (1) ◽  
pp. 117-118
Author(s):  
H. PAUL A. ILLING ◽  
ESTHER S. A. HOUSE
Keyword(s):  
Rat Liver ◽  
High Speed ◽  
High Speed Supernatant

scholarly journals Studies on the meta and para O-sulphation of the catechol compound 3,4-dihydroxybenzoic acid by rat liver sulphotransferase in vitro

1980 ◽  
Vol 191 (1) ◽  
pp. 133-138 ◽  
Author(s):  
E J M Pennings ◽  
G M J Van Kempen
Keyword(s):  
Chemical Synthesis ◽  
Rat Liver ◽  
High Speed ◽  
Helix Pomatia ◽  
Dihydroxybenzoic Acid ◽  
High Speed Supernatant ◽  
Optimal Ph

The enzymic meta and para O-sulphation of 3,4-dihydroxybenzoic acid was investigated in vitro with a dialysed high-speed supernatant from rat liver. The O-sulphated products were identified by comparison with the reference compounds. The chemical synthesis and identification of the reference O-sulphate esters is described in detail. The sulphotransferase activity of the dialysed supernatant from rat liver towards 3,4-dihydroxybenzoic acid was 580 pmol of 3-O-sulphate and 120 pmol of 4-O-sulphate formed/min per mg of protein at the optimal pH of 7.4. The meta/para ratio of O-sulphation was independent of pH, time of incubation, concentration of enzyme and presence of dithiothreitol. The O-sulphate esters of 3,4-dihydroxybenzoic acid were found to be good substrates for the arylsulphatase reaction at pH 5.6. The arylsulphatase activity of a dialysed preparation from rat liver was 4.0 nmol of 3-O- and 5.7 nmol of 4-O-sulphate ester hydrolysed/min per mg of protein, respectively. Arylsulphatase from Helix pomatia had an activity of 620 pmol of 3-O-sulphate and of 16.6 nmol of 4-O-sulphate ester hydrolysed/min per unit (mumol/h) of sulphatase.

scholarly journals Purification of membrane-bound galactosyltransferase from rat liver microsomal fractions

1977 ◽  
Vol 164 (3) ◽  
pp. 541-547 ◽  
Author(s):  
Ian H. Fraser ◽  
Sailen Mookerjea
Keyword(s):  
Molecular Weight ◽  
Rat Liver ◽  
Column Chromatography ◽  
High Speed ◽  
Affinity Column ◽  
High Molecular Weight Fraction ◽  
Serum Enzyme ◽  
Membrane Enzyme ◽  
Membrane Bound ◽  
High Speed Supernatant

1. Rat liver microsomal preparations incubated in 1% Triton X-100 at 37°C for 1h released about 60% of the membrane-bound UDP-galactose–glycoprotein galactosyltransferase (EC 2.4.1.22) into a high-speed supernatant. The supernatant galactosyltransferase which was solubilized but not purified by this treatment had a higher molecular weight than the serum enzyme as shown by Sephadex G-100 column chromatography. 2. The galactosyltransferase present in the high-speed supernatant was purified 680-fold by an affinity-column-chromatographic technique by using a column of activated Sepharose 4B coupled with α-lactalbumin. The galactosyltransferase ran as a single band on polyacrylamide gels and contained no sialyltransferase, N-acetylglucosaminyltransferase or UDP-galactose pyrophosphatase activities. 3. The purified membrane enzyme had properties similar to serum galactosyltransferase. It had an absolute requirement for Mn2+ that could not be replaced by Ca2+, Mg2+, Zn2+ or Co2+, and was active over a wide pH range (6–8) with a pH optimum of 6.5. The apparent Km for UDP-galactose was 10.8μm. The protein α-lactalbumin modified the enzyme to a lactose synthetase by increasing substrate specificity for glucose in preference to N-acetylglucosamine and fetuin depleted of sialic acid and galactose. 4. The molecular weight of the membrane enzyme was 65000–70000, similar to that of the purified serum enzyme. Amino acid analyses of the two proteins were similar but not identical. 5. Sephadex G-100 column chromatography of the purified membrane enzyme showed a small peak (2–5%) of higher molecular weight than the purified serum enzyme. Inclusion of 1mm-ε-aminohexanoic acid in the isolation procedures increased this peak to as much as 30% of the total enzyme recovered. Increasing the ε-aminohexanoic acid concentration to 100mm resulted in no further increase in this high-molecular-weight fraction.

scholarly journals Studies on the purification and properties of UDP-galactose glycoprotein galactosyltransferase from rat liver and serum

1976 ◽  
Vol 156 (2) ◽  
pp. 347-355 ◽  
Author(s):  
I H Fraser ◽  
S Mookerjea
Keyword(s):  
Molecular Weight ◽  
Rat Liver ◽  
High Speed ◽  
Broad Band ◽  
Chromatographic Technique ◽  
Appreciable Effect ◽  
Serum Enzyme ◽  
Membrane Bound ◽  
Triton X ◽  
High Speed Supernatant

1. Rat liver microsomal preparations incubated with 200mM-NaCl at either 0 or 30 degrees C released about 20-30% of the membrane-bound UDP-galactose-glycoprotein galactosyl-transferase (EC 2.4.1.22) into a ‘high-speed’ supernatant. The ‘high-speed’ supernatant was designated the ‘saline wash’ and the galactosyltransferase released into this fraction required Triton X-100 for activation. It was purified sixfold by chromatography on Sephadex G-200, and appeared to have a higher molecular weight than the soluble serum enzyme. 2. Rat serum galactosyltransferase was purified 6000-7000-fold by an affinity-chromatographic technique using a column of activated Sepharose 4B coupled with α-lactalbumin. The purified enzyme ran as a single broad band on polacrylamide gels and contained no sialytransferase, N-acetylglucosaminyltransferase and UDP-galactose pyrophosphatase activities. 3. The highly purified enzyme had properties similar to those of both soluble and membrane-bound galactosyltransferase. It required 0.1% Triton X-100 for stabilization, but lost activity on freezing. The enzyme had an absolute requirement for Mn2+, not replaceable by Ca2+, Mg2+, Zn2+ or Co2+. It was active over a wide pH range (6-8) and had a pH optimum of 6.8. The apparent Km for UDP-galactose was 12.5 × 10(-6) M. α-Lactalbumin had no appreciable effect on UDP-galactose-glycoprotein galactosyltransferase, but it increased the specificity for glucose rather than for N-acetylglucosamine, thus modifying the enzyme to a lactose synthetase. 4. The possibility of a conversion of higher-molecular-weight liver enzyme into soluble serum enzyme is discussed, especially in relation to the elevated activities of this and other glycosyltransferases in patients with liver diseases.

scholarly journals STUDIES ON ISOLATED CELL COMPONENTS

1951 ◽  
Vol 34 (5) ◽  
pp. 647-656 ◽  
Author(s):  
Norman G. Anderson ◽  
Karl M. Wilbur
Keyword(s):  
Rat Liver ◽  
High Speed ◽  
Sucrose Solution ◽  
Anticoagulant Activity ◽  
Liver Homogenate ◽  
Viscosity Increase ◽  
Cell Components ◽  
Liver Homogenates ◽  
Concentration Threshold ◽  
Isolated Cell

1. The addition of heparin to rat liver, kidney, or brain nuclei has been found to bring about the release of a gel. Chemical analysis and histochemical studies on whole homogenates and isolated nuclei demonstrated that the material released by heparin contained desoxyribonucleic acid (DNA) and protein. The action of heparin on nuclei is interpreted as the result of a combination with the basic proteins of the nucleus with a consequent displacement of DNA. 2. The addition of heparin to a finely divided dilute liver homogenate prepared in a phosphate-sucrose solution at pH 7.1 brings about a marked increase in viscosity which reaches a maximum in 6 to 8 minutes at 23° and then declines. 3. The concentration threshold for the viscosity effect was 0.1 mg. per 100 mg. fresh rat liver, with further increases in viscosity at higher heparin concentrations. Over a period of several hours a marked decrease in response to heparin was observed in homogenates stored at 0°. 4. Fractionation of the homogenate demonstrated that the viscosity increase was due to the presence of the nuclei alone, other components showing no effect. Microscopic observation showed that the increase in viscosity was associated with the appearance of a clear gel around nuclei treated with heparin. 5. Heparin brought about the release of DNA from the nuclei of incubated rat liver, kidney, and brain homogenates. In some instances over half the DNA is found in the supernatant after high speed centrifugation (20 minutes, 21,000 x g). 6. No correlation was found between anticoagulant activity of heparin preparations and their effectiveness in causing an increase in the viscosity of liver homogenates. Desulfated heparin produced none of the results described here for heparin.

THE EFFECT OF DIPHENYLHYDANTOIN IN VITRO ON THE METABOLISM OF TESTOSTERONE BY RAT LIVER SUBCELLULAR FRACTIONS

1968 ◽  
Vol 57 (4) ◽  
pp. 649-656
Author(s):  
Leon J. Sholiton ◽  
Emile E. Werk ◽  
Joseph MacGee
Keyword(s):  
Rat Liver ◽  
High Speed ◽  
Metabolite Profile ◽  
Subcellular Fractions ◽  
Polar Metabolite ◽  
Metabolite Production ◽  
Liver Slices ◽  
High Speed Supernatant

ABSTRACT The effect of diphenylhydantoin (DPH) in vitro on the »metabolite profile« of testosterone-4-14C has been studied utilizing incubates of high speed supernatant and microsomal fractions of rat liver. In contrast to earlier experiments with rat liver slices, in which a standard amount of DPH resulted in increased non-polar metabolite production, in the present experiments, when separate subcellular fractions are incubated, no such DPH effect can be detected under the conditions utilized. When certain mixtures of supernatant and microsomal fractions are incubated, however, there is evidence that the DPH augmentation of non-polar metabolite production can again be manifest. An explanation for these results can only be speculated upon at present.

Cholesterol metabolism and vitamin B6. I. Hepatic cholesterogenesis and pyridoxine deficiency

1969 ◽  
Vol 47 (6) ◽  
pp. 631-635 ◽  
Author(s):  
P. J. Lupien ◽  
C. M. Hinse ◽  
M. Avery
Keyword(s):  
Rat Liver ◽  
High Speed ◽  
Cholesterol Metabolism ◽  
Reductase Activity ◽  
Liver Microsomes ◽  
Liver Homogenate ◽  
Supernatant Fraction ◽  
Liver Cholesterol ◽  
Pyridoxine Deficiency ◽  
Hepatic Cholesterogenesis

Hepatic cholesterogenesis was studied in pair-fed and pyridoxine-deficient rats as well as in rat liver homogenate systems. Crossover of various subcellular components from pair-fed homogenates into pyridoxine-deficient homogenate systems and vice versa was also done.On 8 weeks of pyridoxine deficiency, acetate-14C incorporation rates into liver cholesterol increased by a factor of approximately 10. The same phenomenon was observed with the total liver homogenate systems.Pyridoxine deficiency does not appear to affect HMG-CoA reductase activity of pyridoxine-deficient liver microsomes sufficiently to explain the rapid acetate-1-14C incorporation rates in this same tissue. The activating system(s) responsible for the 10-fold increase in acetate-14C incorporation rates into pyridoxine-deficient rat liver cholesterol appears to be located in the high-speed supernatant fraction. Other subcellular components such as lysosomes and mitochondria are probably implicated to some extent in this phenomenon. The results indicate that vitamin B6 is necessary for normal hepatic cholesterogenesis in the rat.The significance of these findings and the possible relationship between these factors are discussed.

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