scholarly journals Characteristics of the rat liver microsomal enzyme system converting cholecalciferol into 25-hydroxycholecalciferol. Evidence for the participation of cytochrome p-450

1979 ◽  
Vol 184 (3) ◽  
pp. 491-499 ◽  
Author(s):  
T C Madhok ◽  
H F DeLuca
Keyword(s):  
Vitamin D ◽  
White Light ◽  
Microsomal Fraction ◽  
Enzyme System ◽  
Initial Reaction ◽  
Microsomal Enzyme ◽  
Reaction Velocity ◽  
Calcium And Phosphorus ◽  
Cytochrome P 450 ◽  
25 Hydroxycholecalciferol

Properties of the rat hepatic cholecalciferol 25-hydroxylase have been studied. An assay system has been developed in which 25-hydroxycholecalciferol production is linear for at least 2h in both homogenates and microsomal fraction. Furthermore, the initial reaction velocity is linearly related to the amount of liver tissue or microsomal fraction. This enzyme system also metabolizes an analogue of cholecalciferol, namely dihydrotachysterol 3, into 25-hydroxydihydrotachysterol 3. The 25-hydroxylase is in the microsomal fraction and not in mitochondria. It has a Km of 44 nM for cholecalciferol and 360 nM for dihydrotachysterol 3. Its activity is not altered by dietary concentrations of calcium and phosphorus. Vitamin D-deficient rats have higher activities of the hepatic 25-hydroxylase than those receiving 25 ng of cholecalciferol daily. The 25-hydroxylase is inhibited by metyrapone. An atmosphere of CO/O2 (9:1, v/v) inhibits the reaction by 87%. This inhibition is partially reversed by white light. Additionally, cholecalciferol and 25-hydroxycholecalciferol competitively inhibit aminopyrine demethylase. These results support the idea that the cholecalciferol 25-hydroxylase is a cytochrome P-450-dependent mono-oxygenase.

INTERMICROSOMAL DISTRIBUTION OF AROMATIZING ENZYME SYSTEM IN EQUINE TESTICULAR TISSUE

1973 ◽  
Vol 72 (2) ◽  
pp. 366-375 ◽  
Author(s):  
Ran Oh ◽  
Bun-ichi Tamaoki
Keyword(s):  
Microsomal Fraction ◽  
Density Gradient Centrifugation ◽  
Enzyme System ◽  
Gradient Centrifugation ◽  
Electron Microscopic Examination ◽  
Sucrose Density Gradient ◽  
Testicular Tissue ◽  
Electron Microscopic ◽  
Sucrose Density Gradient Centrifugation ◽  
Cytochrome P 450

ABSTRACT The microsomal fraction (10 000–105 000 × g precipitate) of equine testes was fractionated into the smooth- and the rough-surfaced microsomal subfractions by a sucrose density-gradient centrifugation in the presence of CsCl. The validity of this fractionating procedure was confirmed by electron microscopic examination and also by chemical analysis of the RNA contents in these subfractions. The aromatizing enzyme system (19-hydroxylase and aromatase) which was concentrated in the microsomal fractions among the organellae was found to be localized in the smoothsurfaced microsomal fraction. The cytochrome P-450 which was also involved in the process of enzymatic aromatization was detected exclusively in the smooth-surfaced microsomal fraction. The distribution of the aromatizing system between the two microsomal subfractions of equine testes was discussed in comparison with that in human full term placentae.

scholarly journals Phospholipid requirement for 2-acetamidofluorene N-and ring-hydroxylation by hamster liver microsomal cytochrome P-450 enzyme system

1977 ◽  
Vol 168 (3) ◽  
pp. 571-574 ◽  
Author(s):  
P D Lotlikar ◽  
E N Dwyer ◽  
W J Baldy ◽  
J Nyce
Keyword(s):  
Total Lipid ◽  
Lipid Content ◽  
Microsomal Fraction ◽  
Enzyme System ◽  
Total Lipid Content ◽  
Appreciable Effect ◽  
Microsomal Cytochrome ◽  
Lower Capacity ◽  
Cytochrome P 450 ◽  
Ring Hydroxylation

A phospholipid requirement of 2-acetamidofluorene N- and ring-hydroxylation was investigated with partially delipidated microsomal fraction from livers of 3-methylcholanthrene-pretreated hamsters. Butan-1-ol extraction of microsomal fraction removed 90% of the total lipid content without any appreciable effect on microsomal proteins. Such extracted microsomal fractions had much lower capacity to N- and ring-hydroxylate 2-acetamidofluorene: 25 and 44% of control respectively. Addition of butan-1-ol-extracted total lipid restored both oxidations to some extent, whereas addition of phosphatidylcholine fraction restored both oxidations almost completely. Addition of synthetic phospholipid, dilauroyl phosphatidylcholine, restored both oxidations to a large extent, whereas synthetic dipalmitoyl or distearoyl phosphatidylcholine was ineffective in restoring these oxidations.

Intestinal CaBP: a new quantitive index of vitamin D deficiency in the rat

1975 ◽  
Vol 229 (3) ◽  
pp. 689-694 ◽  
Author(s):  
F Bronner ◽  
T Freund
Keyword(s):  
Vitamin D ◽  
Vitamin D Deficiency ◽  
Calcium Intake ◽  
Calcium Binding ◽  
Plasma Calcium ◽  
Vitamin D Status ◽  
Calcium And Phosphorus ◽  
Dose Dependent Increase ◽  
Dose Dependent ◽  
25 Hydroxycholecalciferol

Rats raised from weaning on regiments adequate in calcium and phosphorus but deficient in vitamin D will have no detectable intestinal calcium-binding proteins (CaBP), whether or not they show other signs of vitamin D deficiency, such as hypocalcemia. When hypocalcemic, vitamin D-deficient animals were treated with 25-hydroxycholecalciferol, a vitamin D metabolite, they showed a dose-dependent increase in plasma calcium and CaBP; both responses can be described by a single linear relationship, which appears to apply whether the metabolite is 25-hydroxycholecalciferol or dihydrotachysterol. Since vitamin D status is only one determinant of plasma calcium, whereas CaBP (or its expression) appears to depend on vitamin D quantitatively, CaBP may be used as an index of vitamin D status, provided calcium intake is controlled.

Sign in / Sign up

Export Citation Format

Share Document