Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UMLS:C0027960 (mole)
 21,279 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Bilirubin diglucuronide (BDG) may be formed in vitro by microsomal UDP glucuronosyl transferase (EC 2.4.1.17)-mediated transfer of a second mole of glucuronic acid from UDP-glucuronic acid, or by dismutation of bilirubin monoglucuronide (BMG) to BDG and unconjugated bilirubin, catalyzed by an enzyme (EC 2.4.1.95) that is concentrated in plasma membrane-enriched fractions of rat liver. To evaluate the role of these two enzymatic mechanisms in vivo, [(3)H]bilirubin mono-[(14)C]glucuronide was biosynthesized, purified by thin-layer chromatography, and tracer doses were infused intravenously in homozygous Gunn (UDP glucuronyl transferase-deficient) rats or Wistar rats. Bilirubin conjugates in bile were separated by high-pressure liquid chromatography and (3)H and (14)C were quantitated. In Gunn rats, the (14)C:(3)H ratio in BDG excreted in bile was twice the ratio in injected BMG. In Wistar rats the (14)C:(3)H ratio in biliary BDG was 1.25 +/- 0.06 (mean +/- SEM) times the ratio in injected BMG. When double labeled BMG was injected in Wistar rats after injection of excess unlabeled unconjugated bilirubin (1.7 mumol), the (14)C:(3)H ratio in BDG excreted in bile was identical to the ratio in injected BMG. Analysis of isomeric composition of bilirubin conjugates after alkaline hydrolysis or alkaline methanolysis indicated that the bile pigments retained the IX(alpha) configuration during these experiments. The results indicate that both enzymatic dismutation and UDP glucuronyl transferase function in vivo in BDG formation, and that dismutation is inhibited by a high intrahepatic concentration of unconjugated bilirubin. This hypothesis was supported by infusion of [(3)H]bilirubin-monoglucuronide in isolated perfused homozygous Gunn rat liver after depletion of intrahepatic bilirubin by perfusion with bovine serum albumin (2.5%), and after bilirubin repletion following perfusion with 0.34 mM bilirubin. From 20 to 25% of injected radioactivity was recovered in BDG in bile in the bilirubin-depleted state; only 8-10% of radioactivity was in BDG in bile after bilirubin repletion. After infusion of [(3)H]bilirubin di-[(14)C]glucuronide in homozygous Gunn rats, 5-7% of the injected pigment was excreted in bile as BMG. The (14)C:(3)H ratio in the injected BDG was 10% greater than the (14)C:(3)H ratio in BMG excreted in bile. These results indicate that in vivo, dismutation rather than partial hydrolysis, is responsible for BMG formation. Incubation of [(3)H]bilirubin, BDG and a rat liver plasma membrane preparation resulted in formation of BMG (3.3 nmol/min per mg protein) indicating that dismutation is also reversible in vitro.
 ...
PMID:Bilirubin diglucuronide formation in intact rats and in isolated Gunn rat liver. 680 Oct 91

The regulation by the cell of subcellular membrane components is dependent on a highly complex balance of nutritional, hormonal and metabolic events. We have characterized the lipid components of the endoplasmic reticulum (ER) of the liver of adrenalectomized (ADX) rats and the response of these membrane components to glucocorticoid administration. Membrane microviscosity as measured by fluorescence depolarization of 1,6-diphenylhexatriene (DPH) was measured and correlated with lipid composition and content of the membranes. In the ADX rat, a significant increase in membrane microviscosity of the smooth endoplasmic reticulum (SER) was observed and this was accompanied by an increase in the cholesterol content/mg protein and a decrease in the phospholipid content/mg protein. A change in the fatty acyl chain composition is observed with a significant increase in the mole percentage of arachidonic acid (20:4) and an accompanying decrease in saturated fatty acids. Within 2-6 hr of dexamethasone administration, a decrease in membrane microviscosity is observed that returns this value to one similar to that for normal control animals. Both the cholesterol and the phospholipid contents/mg protein are likewise restored to levels similar to that for control animals beginning at the 2-hr time point. The arachidonic acid and saturated fatty acid content of the constituent phospholipids do not begin to return to values similar to those for control animals until 6 hr after dexamethasone administration. From these experiments, we can conclude that glucocorticoids play a significant regulatory role in determining the lipid properties of rat hepatic microsomal membranes.
 ...
PMID:Dynamic lipid changes in rapidly proliferating hepatic smooth endoplasmic reticulum during acute dexamethasone treatment of adrenalectomized rats. 721 73

Ethylmorphine administration to male rats during 3 weeks markedly decreased the cytochrome P-450 content and the rate of oxidation of ethylmorphine, hexobarbital and testosterone in liver microsomes. The decrease of the substrate oxidation rate was correlated with the decrease in the maximal spectral changes induced by the substrate binding to cytochrome P-450. Calculation of the substrate oxidation and binding data per mole of cytochrome P-450 showed that considerable changes were observed in the case of testosterone only. A combined administration of testosterone and ethylmorphine did not prevent the decrease of the substrate oxidation. It was assumed that a prolonged administration of ethylmorphine causes a decline of the form of cytochrome P-450 involved in testosterone metabolism and that testosterone does not directly control the content and activity of microsomal hydroxylases.
 ...
PMID:[Changes in testosterone oxidation in rat liver microsomes after prolonged administration of ethylmorphine]. 724 89

Phospholipid desaturase activity of rat liver microsomes can be varied by dietary manipulation: rats starved and refed a "fat-free" diet have two to three times the activity of normal controls whereas starved rats have no detectable activity. These changes were accompanied by changes in fatty acid composition of the microsomal phospholipids, resulting in a double bond: saturated fatty acid mole ratio (moles double bonds per mole saturated fatty acid) of 5.7 in control and starved rats and 4.7 in starved-refed rats. Fluorescence polarization ratio P (I parallel/I perpendicular to x instrument correction factor) of cis- and trans-parinaric acid (PnA) showed no significant differences in physical state of the three microsomal preparations. However, the isolated microsomal phospholipids with trans-PnA as probe showed differences in the temperature at which onset of a change in polarization ratio occurred (starved-refed greater than normal greater than starved rats). With the cis-PnA as probe, the polarization ratio showed no change in the range 10-40 degrees C but was significantly higher (1.8) in starved-refed rats than in normal and starved rats (1.6 in both cases). These data indicate that the microsomal phospholipids of starved-refed animals were in a less fluid state than those from control and starved rats and that this decrease in fluidity was correlated with an increase in phospholipid desaturase activity.
 ...
PMID:Fluorescence polarization studies of rat liver microsomes with altered phospholipid desaturase activities. 745 88

The endogenous biosynthesis of nitric oxide (NO) is increased during gestation. To begin our investigation of a possible tissue source (or sources), we examined the placenta. We postulated that analogous to the endothelium of blood vessels, the syncytiotrophoblast (STr) cell layer that lines the intervillous blood space of the human placenta would express NO synthase. Our results show that human placental villi express a calcium- and calmodulin-sensitive form of NO synthase, located mainly in the microsomal cell fraction. By in situ hybridization using a riboprobe generated from human endothelial NO synthase cDNA, we observe NO synthase mRNA expression in STr. The STr also shows NADPH-diaphorase staining, indicating the presence of NO synthase, and most likely other flavin-containing enzymes involved in sex steroid metabolism. NO synthase activity was also detected in the villi of a complete mole placenta (which lacks fetal vessels), further supporting a trophoblastic origin. Our findings suggest a previously unrecognized role for STr-derived NO in placental function.
 ...
PMID:Expression of nitric oxide synthase by syncytiotrophoblast in human placental villi. 769 71

The delta 6-desaturase system was isolated from rat liver microsomes by high hydrostatic pressure (1500 bars) and the enzyme components were then separated by size chromatography. The lipids extracted by organic solvents from the pressure shed fractions were phosphatidylcholine (PC) and cholesterol at a mole ratio of 4:1. The acyl chains of the shed PC were 56% saturated and 21% polyunsaturated resulting predominantly from 13% higher and 15% lower contents of palmitic and arachidonic acid, respectively, as compared to those of microsomal PC. The weight ratio of phospholipids to protein in the shed desaturase fraction was 0.2 which corresponds to an average of 31 phospholipid molecules around each desaturase molecule. Differential scanning calorimetry of the lipids associated with the desaturase system showed a phase transition at 41 degrees C. Fluorescence anisotropy studies of the desaturase surrounding lipids indicated the same transition point. We concluded that the delta 6-desaturase has an associated lipid surrounding of PC and cholesterol at an approx. 4:1 mole ratio that constitutes a gel phase at physiological temperature. We suggest that this state is essential for optimal desaturase activity and that the specific acyl chains of the lipid annulus provide a regulatory sensor of the delta 6-desaturase activity.
 ...
PMID:Characterization of the lipid surrounding the delta 6-desaturase of rat liver microsomes. 774 50

A multiple measurement system for assessing sarcoplasmic reticulum (SR) Ca(++)-ATPase activity and Ca(++)-uptake was used to examine the effects of SR fractionation and quick freezing on rat white (WG) and red (RG) gastrocnemius muscle. In vitro measurements were performed on whole muscle homogenates (HOM) and crude microsomal fractions (CM) enriched in SR vesicles before and after quick freezing in liquid nitrogen. Isolation of the CM fraction resulted in protein yields of 0.96 +/- 0.1 and 0.99 +/- 0.1 mg/g in WG and RG, respectively. The percent Ca(++)-ATPase recovery for CM compared to HOM was 14.5% (WG) and 10.1% (RG). SR Ca(++)-activated Ca(++)-ATPase activity was not affected by quick freezing of HOM or CM, but basal ATPase was reduced (P < 0.05) in frozen HOM (5.12 +/- 0.18-3.98 +/- 0.20 mole/g tissue/min in WG and from 5.39 +/- 0.20-4.48 +/- 0.24 mumole/g tissue/min in RG). Ca(++)-uptake was measured at a range of physiological free [Ca++] using the Ca++ fluorescent dye Indo-1. Maximum Ca(++)-uptake rates when corrected for initial [Ca++]f were not altered in HOM or CM by quick freezing but uptake between 300 and 400nM free Ca++ was reduced (P < 0.05) in quick frozen HOM (1.30 +/- 0.1-0.66 +/- 0.1 mumole/g tissue/min in WG and 1.04 +/- 0.2-0.60 +/- 0.1 mumole/g tissue/min in RG). Linear correlations between Ca(++)-uptake and Ca(++)-ATPase activity measured in the presence of the Ca++ ionophore A23187 were r = +0.25, (P < 0.05) and r = +0.74 (P < 0.05) in HOM and CM preparations, respectively, and were not altered by freezing. The linear relationships between HOM and CM maximum Ca(++)-uptake (r = +0.44, P < 0.05) and between HOM and CM Ca(++)-ATPase activity (r = +0.34, P < 0.05) were also not altered by tissue freezing. These data suggest that alterations in maximal SR Ca(++)-uptake function and maximal Ca(++)-ATPase activity may be measured in both HOM and CM fractions following freezing and short term storage.
 ...
PMID:Technical considerations for assessing alterations in skeletal muscle sarcoplasmic reticulum Ca(++)-sequestration function in vitro. 785 41

We have characterized a Brassica rapa mutant (WR586) that has low levels of polyunsaturated fatty acids (D.L. Auld et al., 1992, Crop Sci. 32, 657-662). The mutant lacked oleoyl-phosphatidylcholine desaturase (ODS) activity when assayed in 6-day-old seedlings. To further characterize the mutant, the leaf fatty acid composition and major galactolipids and phospholipids were characterized in mutant (WR586) and control (cultivar "Tobin") plants grown at either 26 degrees C/26 degrees C or 10 degrees C/5 degrees C. Fatty acid profiles show significantly higher 18:1 levels in WR586 throughout 12 days of germination. The amount of saturated fatty acids decreased with a concomitant increase of 18:1. Ratios of 18:1/18:2 revealed that WR586 maintains higher mole percent of 18:1 than Tobin at all times and temperature regimes because of a lack of desaturation to 18:2. Values for monogalactosyl-diacylglycerol and digalactosyldiacylglycerol 18:1/18:2 ratios indicate a disparity in the concentration of 18:1 between WR586 and Tobin grown in either temperature during early germination. The phosphatidylcholine and phosphatidylethanolamine 18:1/18:2 ratios were higher in WR586 compared to Tobin and remained higher throughout the 12-day period. In WR586, 18:1 always accumulated to higher levels in the cooler temperature. In Tobin, 18:1 concentrations paralleled 18:2 in both temperature regimes. These results indicate that the lesion in the mutant WR586 resides at the ODS locus, 18:1 synthesis is chilling induced, and the microsomal desaturation pathway is the most prominent in early developing Brassica seedlings.
 ...
PMID:Biochemical characterization of temperature-induced changes in lipid metabolism in a high oleic acid mutant of Brassica rapa. 797

Mitochondrial NADPH-linked aquacobalamin reductase was purified and characterized to clarify its enzymatic properties. The enzyme was purified about 360-fold over rat liver mitochondrial membranes in a yield of 7.5%. The purified enzyme was homogenous in SDS-PAGE. The molecular mass (M(r)) of the enzyme was calculated to be 65 kDa by SDS-PAGE and by Toyopearl HW55 gel filtration, indicating that the enzyme is a monomeric polypeptide with M(r) of 65 kDa. The enzyme was a flavoprotein containing 1 mol of FAD and FMN per mole of the enzyme. The enzyme was specific for NADPH as electron donor and had the ability to reduce cytochrome c (15.4 mumol.min-1 x mg protein-1), potassium ferricyanide (4.9 mumol.min-1 x mg protein-1) and 2,6-dichlorophenolindophenol (16.8 mumol.min-1.mg protein-1) as well as aquacobalamin (6.4 mumol.min-1 x mg protein-1). Although the enzyme immunoreacted with an antibody against NADPH-cytochrome P-450 reductase, which had the activity of the NADPH-linked aquacobalamin reductase in rat liver microsomes, the mitochondrial enzyme and the microsomal enzyme had different enzymological properties.
 ...
PMID:Mitochondrial NADPH-linked aquacobalamin reductase is distinct from the NADPH-linked enzyme from microsomal membranes in rat liver. 822 2

Oxidation of tienilic acid (TA) by microsomes of yeast expressing two closely related human liver cytochrome P-450s (P450), P450 2C9 and 2C10, led to catalysis-dependent loss of activity of these P450s. Under identical conditions, oxidation of a tienilic acid isomer (TAI) failed to give any P450 inactivation. The loss of P450 activity during TA oxidation was concomitant with product (5-hydroxytienilic acid, 5-OHTA) formation, showed pseudo-first-order and saturation kinetics, and was inhibited by an alternative substrate, tolbutamide. Covalent binding of TA metabolites to microsomal proteins occurred in parallel with enzyme inactivation and was partially inhibited by the presence of glutathione in the reaction medium. However, glutathione did not protect P450 enzyme from inactivation. Thus, TA exhibited all of the characteristics of a mechanism-based inactivator for P450 2C9 and 2C10 enzymes. The following kinetic parameters were determined in the case of P450 2C10: t1/2,max = 3.4 min, k(inact) = 3.6 10(-3) s-1, KI = 4.3 microM, k(inact)/KI = 813 L mol-1 s-1, and partition ratio = 11.6. Moreover, a specific covalent binding of 0.9 mol of TA metabolite per mole of P450 2C10 was found to occur before the complete loss of enzyme activity (in incubations performed in the presence of glutathione). A plausible mechanism for P450 2C10 (2C9) inactivation during TA oxidation is proposed. It involves the intermediate formation of an electrophilic thiophene sulfoxide, which may react at position 5 of its thiophene ring either with H2O to give 5-OHTA or with a nucleophilic group of an amino acid residue of the P450 active site, which results in its covalent binding to P450 protein. This alkylation and inactivation of P450 2C9 (2C10) by TA could be a starting point for the appearance of anti-P450 2C antibodies detected in patients treated with TA and suffering from immunoallergic hepatitis.
 ...
PMID:Thiophene derivatives as new mechanism-based inhibitors of cytochromes P-450: inactivation of yeast-expressed human liver cytochrome P-450 2C9 by tienilic acid. 828 35


<< Previous 1 2 3 4 5 6 Next >>