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1.
Vitamin K dependent carboxylation of glutamate residues to gamma-carboxyglutamate was demonstrated in proteins of spleen and testes microsomes. The rate of carboxylation in spleen microsomes was 0.9% and testes 3% of that in liver microsomes per milligram of microsomal protein. For comparison the rates of carboxylation in lung and kidney microsomes were 17 and 8%, respectively, of the rate in liver microsomes. The high rate in liver microsomes may be due to a high carboxylase level as indicated by the high rate of peptide carboxylation in liver microsomes. Protein carboxylation in liver microsomes was linear for only 15 min while carboxylation in microsomes from extrahepatic tissue persisted much longer so that the total protein carboxylation in lung microsomes was 60%, kidney 18%, testes 12%, and spleen 9% of that occurring in liver microsomes. Protein carboxylation was higher in microsomes from extrahepatic tissues of rats fed a vitamin K deficient diet as compared to animals fed a vitamin K sufficient diet. Protein carboxylation in microsomes from extrahepatic tissues was greatly stimulated by manganese ions and was dependent upon the addition of dithioerythritol. NADH could partially replace the dithiol in spleen, testes, and lung, but NADH-dependent carboxylation was relatively low in kidney and liver microsomes. Dithiol-dependent carboxylation was completely blocked by 10 microM warfarin, but NADH-dependent carboxylation was only slightly inhibited by 100 microM warfarin. Menaquinone-3 was much more active than vitamin K1 in driving carboxylation. Solubilized microsomes catalyzed the carboxylation of glutamate residues to gamma-carboxyglutamate in a pentapeptide Phe-Leu-Glu-Glu-Leu. The rate of carboxylation in lung microsomes was 22%, testes 3.3%, kidney 1.9%, and spleen 1.6% of the rate in liver microsomes. 相似文献
2.
9-Hydroxyellipticine (9-OHE), a potent inhibitor of rat liver monooxygenase activities, binds to the various forms of partially purified lung cytochromes P-450 from untreated and 3-methylcholanthrene (3-MC)-treated rabbits. The spectral data (lambda max: 428 nm (ox.), 447 nm (red.), Ks: 10 microM and 5 muM for cytochrome I and cytochrome II from 3-MC-treated rabbits respectively) resemble those obtained with cytochrome P-450 purified from liver of Aroclor 1254-pretreated rats (lambda max: 428 nm (ox.), 445 nm (red.), Ks: 8 microM). 9-OHE has been shown to inhibit the benzo[a]pyrene hydroxylase activity of rat and rabbit lung microsomes. The inhibitory effect was higher towards the 3-MC-induced lung microsomes than with the control microsomes. However, the lung microsomes, as well as the liver microsomes of rabbits were less sensitive to inhibition by 9-OHE than the corresponding microsomes from rats. These results suggest that rabbit and rat cytochromes P-450 have subtle structural differences. 相似文献
3.
The effects of dietary copper and zinc on free radical production in lung and liver microsomes were studied in male weanling rats. The rats were fed for 6 weeks on one of seven diets, with different copper and zinc concentrations representing low, adequate, and high dietary levels of copper and low and adequate levels of zinc. Rats were put on diets arranged in a 3 X 2 factorial design with copper and zinc supplementations of 0, 15, and 500 mg/kg and 0.5 or 100 mg/kg, respectively. The low copper diet depressed copper levels in both the lungs and liver, although zinc levels were unchanged in rats on the low zinc diets. Endogenous carbon-centered lipid radical production in microsomes induced by NADPH was measured using spin-trapping techniques. The low zinc diets increased free radical production in lung microsomes but not in liver microsomes. No change in free radical production was observed in lung or liver microsomes obtained from rats on low copper diets. The data indicate that endogenous free radical production is increased in lung microsomes as a function of dietary zinc deficiency but is not influenced by copper status. 相似文献
4.
D E Williams H H Meyer M S Dutchuk 《Comparative biochemistry and physiology. B, Comparative biochemistry》1989,93(2):465-470
1. Flavin-containing monooxygenase (FMO) in pulmonary and hepatic microsomes from sheep was analyzed by western blotting by probing with antibodies raised against FMO purified from rabbit lung and pig liver. 2. Pulmonary microsomes from sheep contain a single major protein which cross-reacts with the antibody to rabbit lung FMO, but no band can be observed when probed with the antibody to the pig liver enzyme. Likewise, sheep liver microsomes contain a protein which cross-reacts with the antibody to pig liver FMO, but no significant staining is observed following incubation with antibody to the lung enzyme. 3. Sheep pulmonary and hepatic microsomal FMO also display a difference in activity toward chlorpromazine and n-dodecylamine. 4. Preliminary evidence suggests that sheep FMO may be induced (liver) or repressed (lung) during pregnancy. 5. Sheep are similar to rodents (rat, mouse, guinea pig, hamster and rabbit) in having distinct forms of pulmonary and hepatic FMO. The immunochemical and catalytic difference between sheep liver and lung FMO is similar to that of rabbit. 相似文献
5.
Ethylmorphine N-demethylase activity of the sheep liver and lung microsomes was reconstituted in the presence of solubilized microsomal cytochrome P-450, NADPH-cytochrome c reductase and synthetic lipid, phosphatidylcholine dilauroyl. The Km of the lung microsomal ethylmorphine N-demethylase was calculated to be 4.84 mM ethylmorphine from its Lineweaver-Burk graph and lung enzyme was inhibited by its substrate, ethylmorphine, when its concn was 25 mM and above, reaching to 67% inhibition at 50 mM concn. The Lineweaver-Burk and Eadie-Hofstee plots of the liver enzyme were found to be curvilinear. From these graphs, two different Km values were calculated for the liver enzyme as 4.17 mM and 0.40 mM ethylmorphine. Ethylmorphine N-demethylase activities of both liver and lung microsomes were inhibited by NiCl2, CdCl2 and ZnSO4. Ethylalcohol inhibited N-demethylation of ethylmorphine in lung and liver microsomes. Acetone (5%) slightly enhanced the N-demethylase activity of the liver enzyme, whereas 5% acetone completely inhibited the lung enzyme. Phenylmethylsulfonyl fluoride at 0.10 mM and 0.25 mM concn had no effect on liver enzyme activity, while at these concns, it inhibited the activity of the lung enzyme by about 35%. 相似文献
6.
Harumi Okuyama Kazuyo Yamada Takaki Miyagawa Masahiro Suzuki Renuka Prasad William E.M. Lands 《Archives of biochemistry and biophysics》1983,221(1):99-107
An enzymatic basis for the formation of pulmonary surfactant lipids in rat has been presented. The free fatty acid pools in lung and liver consisted mainly of palmitic, stearic, oleic, and arachidonic acids with relatively less polyunsaturated fatty acids in lung than in liver. The acyl chain specificities of the acyl-CoA synthetase systems in lung and liver microsomes were similar in that most of fatty acids found in the free fatty acid pools were effectively activated by both systems. The acyl-CoA pools had compositions significantly different from those of the free fatty acid pools in lung and liver with relatively more stearate and less polyunsaturated fatty acids. The lung acyl-CoA pool contained mainly palmitate (29%), stearate (31%), and oleate (22%) with very little polyunsaturated acyl-CoAs to compete for esterification. The use of an equimolar mixture of palmitoyl-CoA and arachidonoyl-CoA to acylate the endogenous monoacyl-glycerophosphocholine isomers in the lung microsomes yielded both the 2-palmitate and 2-arachidonate diacyl forms, whereas the major products formed by liver microsomes were the 2-arachidonate and 1-palmitate forms. These results indicate that the 1-acyl isomer is the major monoacyl-glycerophosphocholine species serving as substrate in lung microsomes, whereas both 1-acyl and 2-acyl isomers are present in liver microsomes. Thus, the enrichment of saturated and oligoenoic acids in the acyl-CoA pool combined with the predominance of the 1-acyl isomer in the acyl acceptor pool and the relatively higher selectivity for palmitoyl-CoA by the 1-acyl-GPC acyltransferase activity of lung constitute an important basis for attributing some of the formation of pulmonary surfactant lipids in rats to acyltransferase action. 相似文献
7.
Lung microsomal membranes that contain the redox active components associated with the mixed-function oxidase system can be peroxidized in vitro. To investigate the characteristics of rat lung microsomal lipid peroxidation, we performed experiments using a variety of peroxidation initiators and microsomes obtained from normal and vitamin E-deficient rats. We found that lung microsomes obtained from normal rats are peroxidized much less than liver microsomes obtained from the same animals. Only initiation systems using very high concentrations of ferrous iron produced any significant peroxidation of normal rat lung microsomes. Lung microsomes obtained from vitamin E-deficient rats were found to be much more susceptible to peroxidation. Glutathione (GSH) was effective in inhibiting peroxidation when lung microsomes from normal rats were peroxidized. GSH was not effective in decreasing peroxidation when microsomes from vitamin E-deficient rats were peroxidized in the same system. We conclude that both GSH and vitamin E protect lung microsomal membranes from peroxidation. Glutathione protection appears to be related to the presence of a sulfhydryl group. 相似文献
8.
Rabbit lung and liver microsomes were subjected to three procedures which decreased NADPH cytochrome c reductase activity; flavoprotein antibody, trypsin and subtilisin digestion. The effects on benzphetamine and p-nitroanisole demethylation and amine metabolic-intermediate complex formation were investigated. In general, the proteolytic digestion had a greater inhibitory effect on oxidation reactions for a given loss of NADPH cytochrome c reductase activity than did flavoprotein antibody; and of the two proteases, subtilisin, which also diminishes the cytochrome b5 reduction pathway, had a greater inhibitory effect than trypsin. Subtilisin digestion had similar effects in both liver and lung microsomes; a loss of flavoprotein without a loss of cytochrome P-450; but whereas all three oxidative reactions decreased in unison as the flavoprotein was lost in the liver, benzphetamine demethylation was less susceptible to flavoprotein depletion than the other two reactions in lung microsomes. With trypsin digestion flavoprotein was removed without loss of cytochrome P-450 only in lung microsomes; in liver microsomes the cytochrome P-450 was susceptible to tryptic degradation. In lung microsomes, benzphetamine and p-nitroanisole demethylations were less susceptible to flavoprotein loss than metabolic-intermediate complex formation. 相似文献
9.
Piergiacomi VA Palacios A Catalá A 《Prostaglandins, leukotrienes, and essential fatty acids》2001,64(2):75-80
The effect of retinyl palmitate on the polyunsaturated fatty-acid composition, chemiluminescence and peroxidizability index of microsomes and mitochondria obtained from rat liver, kidney, brain, lung and heart, was studied. After incubation of microsomes and mitochondria in an ascorbate Fe++ system (120 min at 37 degrees C) it was observed that the total cpm/mg protein originated from light emission: chemiluminescence was lower in liver microsomes, mitochondria and kidney microsomes in the vitamin A group than in the control group. In mitochondria obtained from control rats, the most sensitive fatty acids for peroxidation were arachidonic acid C20:4 n6 in liver and docosahexaenoic acid C22:6 n3 in kidney and brain. In microsomes obtained from control rats, the most sensitive fatty acids for peroxidation were linoleic acid C18:2 n6 and C20:4 n6 in liver and C22:6 n3 in kidney. Changes in the most polyunsaturated fatty acids were not observed in organelles obtained from lung and heart. As a consequence the peroxidizability index, a parameter based on the maximal rate of oxidation of fatty acids, showed significant changes in liver, kidney and brain mitochondria, while in microsomes changes were significant in liver and kidney. These changes were less pronounced in membranes derived from rats receiving vitamin A. Our results confirm and extend previous observations that indicated that vitamin A may act as an antioxidant protecting membranes from deleterious effects. 相似文献
10.
Synthesis of phosphatidylglycerol from CDPdiacylglycerol and glycerol 3-phosphate by membranous subcellular fractions of rat lung and liver was optimal when assayed in the presence of bovine serum albumin and Triton X-100. Specific activities of glycerolphosphate phosphatidyltransferase in all membranous subcellular fractions of lung were several times higher than the corresponding fractions from liver. Distribution of this enzyme in subcellular fractions of lung or liver closely parallel the activity of the mitochondrial enzymes monoamine oxidase and succinate cytochrome c reductase. The phosphatidylglycerol-synthesizing activity in microsomes of both lung and liver was a minor fraction of total tissue activity and could be interpreted as due either to contamination with outer mitochondrial membrane or to a small amount of activity innate to microsomes. These results suggest that phosphatidylglycerol, which is believed to be a component of pulmonary surfactant, is synthesized by lung at a rapid rate relative to liver and that the subcellular distribution of its synthesis is similar in both tissues, with mitochondria as the major site. 相似文献
11.
Generalizations derived from experimental studies of enzymic disorganization and stability of membranes of cellular structures (microsomes, mitochondria, lysosomes, cytoplasmic membrane) of various organs and systems (liver, kidney, lung, brain, placenta, gonads, alveolar macrophages and others) due to hepato-, neuro-, gonado-, embryotoxic, mutagenic and carcinogenic effect of chemical pollutants of the environment are presented. Evaluation of the membrane-damaging effect taking into account the stage of labilization of biomembranes (according to severity of manifestation) and compensatory defence reactions at the moleculo-cellular level (alveolar macrophages) is recommended. Classification of biochemical methods (according to priority) is proposed for the evaluation of early functional changes during the examination of the health state of the population in dependence on the effect of environmental factors. 相似文献
12.
Metabolism of benzo(a)pyrene by microsomes from tissues of pregnant and fetal hamsters 总被引:1,自引:0,他引:1
Pretreatment of hamsters with benzo (a) pyrene (BaP) greatly increased the metabolism of BaP by lung microsomes from pregnant hamsters, and had less effect on the metabolism of BaP by liver microsomes. The production of various metabolites of BaP by lung microsomes was increased to different extents: 3-hydroxy-BaP (3-OH-BaP) was one of the major metabolites; the metabolic yields of 9, 10-dihydrodihydroxy-BaP (9, 10-diol) and 7,8-diol were increased more than that of the 4,5-diol. In the case of liver microsomes, only the yields of 9,10-diol and 7,8-diol were increased over the control levels. The presence of cyclohexene oxide in the incubation mixtures decreased the production of the diols. Basal-level enzyme activities in placental, fetal liver, and fetal skin microsomes in metabolizing BaP were very low. Pretreatment of pregnant hamsters with BaP induced BaP-metabolizing enzymes in fetal tissue 2–3 fold. 相似文献
13.
Krishnappa Venkatesh Bonnie Blake Patricia E. Levi Ernest Hodgson 《Journal of biochemical and molecular toxicology》1992,7(3):163-169
The flavin-containing monooxygenase (FMO) was purified from mouse lung microsomes. On SDS-PAGE, the purified enzyme separated as two bands, a major band of 58,000 daltons and a minor band of 59,000 daltons. Antibodies to mouse liver FMO cross-reacted with both bands in the purified preparations, whereas antibodies to rabbit lung FMO cross-reacted only with the major band. In microsomal preparations the major band was recognized by both antibodies, but neither antibody detected the minor band in microsomes. A cDNA encoding the pig liver FMO hybridized with mRNA isolated from mouse liver, kidney, and lung, whereas cDNA encoding the rabbit lung FMO hybridized only with mouse lung and kidney mRNA. Thermal stability studies showed that the FMO preparation purified from mouse lung consisted of a heat-stable and a heat-labile component. The heat-labile component of lung FMO was inhibited competitively by imipramine, whereas the heat-stable component was insensitive to the presence of imipramine. Immunoprecipitation of purified mouse lung FMO with anti-rabbit lung FMO completely removed the protein band reactive to anti-rabbit lung FMO while leaving reactivity to anti-liver FMO. The catalytic and immunochemical differences seen between FMO from rabbit lung and mouse lung appear to result from the expression of at least two forms of FMO in the mouse lung, one similar to the rabbit pulmonary form and one similar to the major mouse liver form of FMO. 相似文献
14.
Zahra Parandoosh Michael R. Franklin 《Biochimica et Biophysica Acta (BBA)/General Subjects》1983,760(3):327-334
Rabbit lung and liver microsomes were subjected to three procedures which decreased NADPH cytochrome c reductase activity; flavoprotein antibody, trypsin and subtilisin digestion. The effects on benzphetamine and p-nitroanisode demethylation and amine metabolic-intermediate complex formation were investigated. In general, the proteolytic digestion had a greater inhibitory effect on oxidation reactions for a given loss of NADPH cytochrome c reductase activity than did flavoprotein antibody; and of the two proteases, subtilisin, which also diminises the cytochrome b5 reduction pathway, had a greater inhibitory effect than trypsin. Subtilisin digestion had similar effects in both liver and lung microsomes; a loss of flavoprotein without a loss of cytochrome P-450; but whereas all three oxidative reactions decreased in unison as the flavoprotein was lost in the liver, benzphetamine demethylation was less susceptible to flavoprotein depletion than the other two reactions in lung microsomes. With trypsin digestion flavoprotein was removed without loss of cytochrome P-450 only in lung microsomes; in liver microsomes the cytochrome P-450 was susceptible to tryptic degradation. In lung microsomes, benzphetamine and p-nitroanisole demethylations were less susceptible to flavoprotein loss than metabolic-intermediate complex formation. 相似文献
15.
Clara cells, alveolar type II cells and pulmonary alveolar macrophages (PAM) were isolated in high yield from rabbit lung. The purity of the cell fractions was 80–90%, 98% and above 99%, respectively. Cytochrome P-450 total content was determined in microsomes from freshly prepared cells. The Clara cells contained significantly more cytochrome P-450 than was found in whole lung microsomes. Furthermore, the cytochrome content of the Clara cells was 2 -fold higher than in the type II cells and 4 -fold higher than in the macrophages. 2-aminofluorene (AF) was the major metabolite in all preparations when intact cells were incubated with 2-acetylaminofuorene (AAF). The PAMs produced AF in the highest rates, while the Clara cells showed the largest rates of cytochrome P-450-dependent, ring hydroxylation of AAF. Mutagenic activation of AAF by isolated lung cells was assayed with a chamber-incubation method. The Clara cells were far more active than the type II cells in this respect, while the macrophages were inactive.Abbreviations AAF
2-acetylaminofluorene
- AF
2-aminofluorene
- DMSO
dimethyl sulfoxide
- NBT
nitro blue tetrazolium
- 7-OH-AAF
7-hydroxy-AAF
- 9-OH-AAF
9-hydroxy-AAF 相似文献
16.
Metabolism of prostaglandins, prostaglandin analogs and thromboxane B2 by lung and liver microsomes from pregnant rabbits 总被引:1,自引:0,他引:1
W S Powell 《Biochimica et biophysica acta》1979,575(3):335-349
Liver microsomes from pregnant rabbits converted prostaglandins F2 alpha, E1, and E2 to their 20-hydroxy metabolites along with smaller amounts of the corresponding 19-hydroxy compounds. Prostaglandins E1 and E2 were also reduced to prostaglandins F1 alpha and F2 alpha, respectively, and prostaglandin E1 was isomerized to 8-isoprostaglandin E1. The above products were also identified after incubation of prostaglandins with liver microsomes from non-pregnant rabbits. In this case, the yield of 20-hydroxy metabolites was much lower. Thromboxane B2 and a number of prostaglandin F2 alpha analogs were also hydroxylated by lung and liver microsomes from pregnant rabbits. The relative rates of hydroxylation by lung microsomes were: prostaglandin E2 approximately prostaglandin F2 alpha approximately 16,16-dimethylprostaglandin F2 alpha approximately 13,14-didehydroprostaglandin F2 alpha greater than thromboxane B2 greater than 15-methylprostaglandin F2 alpha approximately 17-phenyl-18,19,-20-trinorprostaglandin F2 alpha approximately ent-13,14-didehydro-15-epiprostaglandin F2 alpha. Similar results were obtained with liver microsomes except that thromboxane B2 was a relatively poorer substrate for hydroxylation. 相似文献
17.
Pregnant female Wistar rats that received a control (100 ppm Zn) or a Zn-deficient diet (1.5 ppm Zn) from d 0 to 21, or nonpregnant normally fed female rats without or with five daily oral doses of 300 mg/kg salicylic acid were used for the experiments. In isolated mitochondria or microsomes from various maternal and fetal tissues, lipid peroxidation was determined as malondialdehyde formation measured by means of the thiobarbiturate method. Zn deficiency increased lipid peroxidation in mitochondria and microsomes from maternal and fetal liver, maternal kidney, maternal lung microsomes, and fetal lung mitochondria. Lipid peroxidation in fetal microsomes was very low. Zn deficiency produced a further reduction of lipid peroxidation in fetal liver microsomes. Salicylate increased lipid peroxidation in liver mitochondria and microsomes after addition in vitro and after application in vivo. The increase of lipid peroxidation by salicylate may be caused by two mechanisms: an increased cellular Fe uptake that, in turn, can increase lipid peroxidation and chelating Fe, in analogy to the effect of ADP in lipid peroxidation. The latter effect of salicylate is particularly expressed at increased Fe content. 相似文献
18.
C Casals L M Herrero Y Soriano P García-Barreno A M Municio 《Biochemical and biophysical research communications》1985,126(1):551-558
A highly monitorized animal model has been developed for the study of the influence of low oxygen tension on lipid composition, microviscosity and regulation of enzyme activities involved in the phospholipid synthesis of hepatic and pulmonary microsomes. Microviscosity decreased in liver microsomes whereas no difference was shown in that of microsomal membrane core of hypoxemic lung. Nevertheless, phospholipid and cholesterol content of both liver and lung membranes changed significantly. Microsomal membranes of hypoxemic liver increased the unsaturation degree of fatty acids, whereas hypoxemic lung membranes become more saturated, mainly due to the increase of palmitic acid. The adaptive response of lung was confirmed by the high increase of the deacylation-reacylation mechanism. 相似文献
19.
The effect of potassium cyanide on the desaturase activity for stearyl-CoA in microsomes of various mammalian tissues has been investigated. Potassium cyanide inhibited the desaturase activities in microsomes from livers of rat, hen, guinea pig and rat lung, but not the activities in microsomes from rabbit liver, pig thyroid and bovine adrenocortex, It is concluded that the so-called "cyanide-sensitive factor" does not seem to be common to all the desaturase activities of stearyl-CoA in the microsomes of mammalian tissues. 相似文献
20.
Identification of vitamin K-dependent carboxylase activity in lung type II cells but not in lung macrophages. 总被引:3,自引:3,他引:0
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Fluorography of 14C-labelled glutamic acid residues in vitamin K-dependent protein precursors in lung microsomes (microsomal fractions) shows that the lung has several substrates that are not found in the liver. These precursor proteins unique to the lung have apparent molecular masses of 65, 53, 50, 36, 31 and 13 kDa. Type II epithelial cells appear to synthesize most of the vitamin K-dependent proteins in the lung. The 36 and the 31 kDa precursors also found in Type-II-cell microsomes have a similar molecular mass to those of surfactant-associated proteins, and we have previously shown [Rannels, Gallaher, Wallin & Rannels (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 5952-5956] that the 36 kDa protein is one of the precursors for these proteins. Immunoblotting of membrane fragments of Type-II-cell microsomes with plasma prothrombin antibodies identified two prothrombin-like antigens of apparent molecular masses 68 and 65 kDa. This raises the question as to whether Type II cells are also a potential site for synthesis of prothrombin and possibly other vitamin K-dependent clotting factors. Pulmonary macrophages appear to be devoid of vitamin K-dependent carboxylase activity. However, Type II epithelial cells have significant activity, and this activity was unaltered when these cells were maintained in primary culture for 3 days, suggesting that carboxylase activity is expressed in lung alveolar epithelium independently of culture-induced changes in cellular differentiation. Carboxylase activity in Type II cells was enhanced 2-fold when cells were cultured for 24 h in the presence of 50 microM-warfarin. Type II cells, therefore, resemble hepatocytes with regard to their response to coumarin anticoagulant drugs. 相似文献