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1.
CHOLINE METABOLISM AND MEMBRANE FORMATION IN RAT HEPATOMA CELLS GROWN IN SUSPENSION CULTURE : II. Phosphatidylcholine Synthesis during Growth Cycle and Fluctuation of Mitochondrial Density 总被引:4,自引:1,他引:3
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Peter G. W. Plagemann 《The Journal of cell biology》1969,42(3):766-782
The incorporation of methyl-labeled choline into phosphorylcholine and phosphatidylcholine of cellular membranes by Novikoff rat hepatoma cells (line N1S1-67) during growth in suspension culture was investigated. Upon initiation of a fresh culture at 105 cells/ml, the rate of synthesis of phosphorylcholine by the cells was four to five times greater than that of the synthesis of phosphatidylcholine. While the rate of synthesis of the latter remained relatively constant, the rate of phosphorylation of choline decreased progressively during the course of the growth cycle of the culture to 10–20% of the initial rate when the culture reached stationary phase at 3 x 106 cells/ml. The decrease in phosphorylcholine synthesis during the growth cycle was not due to depletion of choline in the medium or a decrease in its concentration, but was correlated with a decrease in choline kinase activity of the cells as measured in cell-free extracts. Newly synthesized phosphatidylcholine was detectable in cells only as an integral part of cellular membranes. Its distribution among various cytoplasmic membrane structures separated by isopycnic centrifugation in sucrose density gradients remained relatively constant during the growth cycle. About 50% was associated with the mitochondria, and the remainder with plasma membrane fragments and other membranous structures with mean densities of about 1.15 and 1.13 g/cm3, respectively. However, the density of the mitochondria increased from about 1.167 g/cm3 in early exponential phase cells to about 1.190 g/cm3 in stationary phase cells. The finding that the density of the entire propulation of mitochondria changed simultaneously and progressively is in agreement with the view that mitochondria grow by addition of phospholipids and structural proteins and increase in number by division. 相似文献
2.
Effect of Mengovirus Replication on Choline Metabolism and Membrane Formation in Novikoff Hepatoma Cells 总被引:6,自引:5,他引:1
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Infection of Novikoff rat hepatoma cells (subline NlSL-67) with mengovirus resulted in a two- to threefold increase in the rate of choline incorporation into membrane phosphatidylcholine at about 3 hr after infection, without affecting the rate of transport of choline into the cell or its phosphorylation. The time course of virus-stimulated phosphatidylcholine synthesis was compared with the time courses of other virus-induced processes during a single cycle of replication. The formation of viral ribonucleic acid (RNA) polymerase and of viral RNA commenced about 1 hr earlier than the virus-stimulated choline incorporation. Further, isopycnic centrifugation of cytoplasmic extracts indicated that the excess of phosphatidylcholine synthesized by infected cells is not located in the membrane structures associated with the viral RNA replication complex, but with structures of a lower density (1.08 to 1.14 g/cc). These membrane structures probably represent the smooth vesicles which accumulate in the cytoplasm of infected cells during the period of increased phosphatidylcholine synthesis between 3 and 5 hr after infection. They are formed with both newly synthesized phosphatidylcholine and phosphatidylcholine present prior to infection. However, concomitant protein synthesis is not required for the stimulated synthesis of membranes; the effect was not inhibited by treating the cells with inhibitors of protein synthesis at 3 hr after infection, although virus production was inhibited about 90% and virus-induced cell degeneration was markedly reduced and delayed. Production of mature virus began normally at about the same time as the stimulation of phosphatidylcholine synthesis. Treatment of infected cells with puromycin at 2 hr, on the other hand, completely inhibited the stimulation of phosphatidylcholine synthesis. 相似文献
3.
Choline metabolism and phosphatidylcholine biosynthesis in cultured rat hepatocytes. 总被引:5,自引:3,他引:2
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1. Adult rat hepatocytes were isolated by collagenase perfusion and were maintained in monolayer culture for 24h. 2. Choline metabolism and phosphatidylcholine biosynthesis were studied in these cells by performing pulse-chase studies at physiological concentrations (1-40 microM) of (Me-3H)-labelled or unlabelled choline in the culture medium. 3. During the 15 min pulse incubation, choline entering the cells was rapidly phosphorylated to phosphocholine or oxidized to betaine. Low concentrations of choline in the medium decreased the relative amount of choline oxidized. 4. During the 3 h chase period, the radioactivity in the phosphocholine pool was transferred to phosphatidylcholine. Very little radioactivity was associated with CDP-choline. These results provide good evidence that the rate-limiting step for phosphatidylcholine biosynthesis in these cultured hepatocytes is the conversion of phosphocholine into CDP-choline. Similar results were obtained for all concentrations of choline in the culture medium. 5. Cellular concentrations of phosphocholine were unaffected by the concentration of choline (1-40 microM) in the medium. 6. The majority of the label associated with betaine was secreted into the culture medium during the chase incubation. 7. From the pulse-chase studies, and the cellular phosphocholine concentrations, it was possible to estimate the rate of phosphatidylcholine biosynthesis (2.2, 2.8, 3.1 and 3.7 nmol/min per g wet weight of cells cultured in 1, 5, 10 and 40 microM-choline respectively for up to 4.25 h). 相似文献
4.
P A Weinhold D A Feldman M M Quade J C Miller R L Brooks 《Biochimica et biophysica acta》1981,665(1):134-144
The sequence of reactions which function to incorporate choline into phosphatidylcholine was investigated in lung from fetuses following premature delivery. The rate of [methyl-14C]choline incorporation by rat lung slices into phosphatidylcholine increases following premature delivery at both 20 and 21 days gestation. The increase in choline incorporation is primarily due to an increased specific activity of phosphorylcholine resulting from a decreased pool size of phosphorylcholine. The decrease in the concentration of phosphorylcholine following premature delivery is apparently caused by an increased activity of cytidylyltransferase which leads to an increase in the conversion of phosphorylcholine to phosphatidylcholine. The total activity of choline kinase, cytidylyltransferase, cholinephosphotransferase and phosphatidate phosphohydrolase did not change significantly. However, the cytidylyltransferase activity in the microsome fraction increased following premature delivery at 20 and 21 days gestation. The amount of cytidylyltransferase in the H form in the cytosol fraction increased following premature delivery at 21 days gestation but not at 20 days gestation. The results are interpreted to indicate that the active form of cytidylyltransferase in lung cells is the membrane-bound enzyme and this form increases following birth resulting in an increased synthesis of phosphatidylcholine. 相似文献
5.
Abstract: The phosphorylcholine concentration of rat brain rises and falls in response to parallel changes in the concentration of circulating choline. A single oral dose of choline chloride (20 mmol/kg) elevated whole-brain concentrations of both choline and phosphorylcholine 5 h after administration; a greater proportion of exogenously administered choline was retained by the brain in its phosphorylated form than as the free arnine. Striatal phosphorylcholine concentrations were elevated within 2 h of choline administration and continued to be significantly greater than control values for up to 34 h after treatment. The response of striatal choline levels to exogenous choline was of shorter duration than that of phosphorylcholine and was correlated with a significant increase in striatal acetylcholine concentrations. The consumption of a choline-free diet for 7 days lowered both serum choline and striatal phosphorylcholine concentrations, but had no effect on striatal choline or acetylcholine. These results suggest that choline kinase is unsaturated by its substrate in vivo and may thus serve to modulate the response of brain choline concentrations to alterations in the supply of circulating choline. 相似文献
6.
Treatment of cultured cells with phospholipase C causes increased rates of hydrolysis of cellular phosphatidylcholine and increased rates of incorporation of choline into phosphatidylcholine. The fate of the diacylglycerol produced by the phospholipase C hydrolysis was examined in two cell lines, Chinese hamster ovary and HeLa. In the former cells, turnover of the glycerol moiety of phosphatidylcholine was not enhanced by phospholipase C treatment, indicating that the phospholipase C-generated diacylglycerol was recycled into new phosphatidylcholine. In HeLa cells, turnover of the glycerol backbone of phosphatidylcholine was enhanced by phospholipase C treatment, and the increased rate of turnover of the glycerol moiety was similar to that of the phosphate moiety. Thus, the fate of diacylglycerol generated at the plasma membrane was demonstrated to differ in these two cell lines. Incorporation of precursors of diacylglycerol into phosphatidylcholine was not enhanced by phospholipase C treatment in either cell line. 相似文献
7.
The incorporation of [methyl-14C]choline into the choline-containing compounds of Ascaris suum muscle and the effects of acetylcholine and its agonists, carbachol and levamisole, on this incorporation were studied. Previous experiments reported a stimulation of phosphatidylcholine (lecithin) metabolism upon the administration of acetylcholine. Acetylcholine administered in vitro to A. suum muscle and body wall preparations resulted in a stimulation of phospholipase C activity that, in turn, produced an increased rate of hydrolysis of phosphatidylcholine to the corresponding diacylglyceride (DAG). The DAG, in turn, may act as a second messenger as it is required for the activation of an A. suum protein kinase C. Evidence presented here is in accordance with this hypothesis. The administration of cholinergics resulted in a stimulation of phosphatidylcholine turnover. Acetylcholine also stimulated isotope incorporation into glycerophosphorylcholine, presumably as a consequence of enhanced phospholipid turnover. These events appear to be associated with the ligand binding to the acetylcholine receptors of the A. suum muscle. Choline kinase activity is suggested in order to maintain the observed high ratio of phosphorylcholine to choline. Findings indicate that in the parasite's muscle phosphatidylcholine metabolism may be linked to receptor-dependent responses and subsequent signal transduction. 相似文献
8.
Jochen Klein rea Köppen Konrad Löffelholz Jorg Schmitthenner 《Journal of neurochemistry》1992,58(3):870-876
The present study is concerned with the uptake and metabolism of choline by the rat brain. Intraperitoneal administration of choline chloride (4-60 mg/kg) caused a dose-dependent elevation of the plasma choline concentration from 11.8 to up to 165.2 microM within 10 min and the reversal of the negative arteriovenous difference (AVD) of choline across the brain to positive values at plasma choline levels of greater than 23 microM. Net choline release and uptake were linearly dependent on the plasma choline level in the physiological range of 10-50 microM, whereas the CSF choline level was significantly increased only at plasma choline levels of greater than 50 microM. The bolus injection of 60 mg/kg of [3H]choline chloride caused the net uptake of greater than 500 nmol/g of choline by the brain as calculated from the AVD, which was reflected in a minor increase of free choline level and a long-lasting increase of brain phosphorylcholine content, which paralleled the uptake curve. Loss of label from phosphorylcholine 30 min to 24 h after choline administration was accompanied by an increase of label in phosphatidylcholine, an indication of a delayed transfer of newly taken-up choline into membrane choline pools. In conclusion, homeostasis of brain choline is maintained by a complex system that interrelates choline net movements into and out of the brain and choline incorporation into and release from phospholipids. 相似文献
9.
Cytidine, as cytidine 5'-diphosphate choline, is a major precursor in the synthesis of phosphatidylcholine in cell membranes. In the present study, we examined the relationships between extracellular levels of cytidine, the conversion of [14C]choline to [14C]phosphatidylcholine, and the net syntheses of phosphatidylcholine and phosphatidylethanolamine by PC12 cells. The rate at which cytidine (as [3H]cytidine) was incorporated into the PC12 cells followed normal Michaelis-Menten kinetics (Km = 5 microM; Vmax = 12 x 10(-3) mmol/mg of protein/min) when the cytidine concentrations in the medium were below 50 microM; at higher concentrations, intracellular [3H]cytidine nucleotide levels increased linearly. Once inside the cell, cytidine was converted mainly into cytidine triphosphate. In pulse-chase experiments, addition of cytidine to the medium caused a time- and dose-dependent increase (by up to 30%) in the incorporation of [14C]choline into membrane [14C]-phosphatidylcholine. When the PC12 cells were supplemented with both cytidine and choline for 14 h, small but significant elevations (p less than 0.05) were observed in their absolute contents of membrane phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine, all increasing by 10-15% relative to their levels in cells incubated with choline alone. Exogenous cytidine, acting via cytidine triphosphate, can thus affect the synthesis and levels of cell membrane phospholipids. 相似文献
10.
《Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism》1980,617(2):225-236
- 1.1. Type II cells, maintained in an organotypic system where in vivo morphologic characteristics are retained, were utilized to study surfactant phospholipid and protein synthesis. Cellular components were separated into a surfactant and a residual fraction by sucrose density centrifugation. The major phospholipid of the surfactant fraction is phosphatidylcholine (70%). Phosphatidylglycerol accounts for 4.0% of the total surfactant phospholipids. Phosphatidylcholine is also the major phospholipid of the residual fraction, constituting 50% of this fraction's phospholipids; phosphatidylglycerol is also present (4.2%).
- 2.2. Type II cells in organotypic culture are capable of metabolizing exogenous glucose. The uptake of glucose from the incubation medium is concentration dependent. Glucose can be utilized by the type II cell for surfactant and residual phosphatidylcholine synthesis. The rate of incorporation of glucose into residual phosphatidylcholine is three times the incorporation rate into surfactant phosphatidylcholine.
- 3.3. Palmitate and choline can also be utilized for surfactant and residual phosphatidylcholine synthesis. Palmitate incorporation into residual phosphatidylcholine is five times greater than the incorporation into surfactant phosphatidylcholine. Surfactant phosphatidylcholine is labelled with choline at a rate one-fourth that of residual phosphatidylcholine. In contrast to glucose and palmitate, choline incorporation into surfactant phosphatidylcholine is linear only after a 15 min lag period. The labelling of residual phosphatidylcholine with choline is biphasic.
- 4.4. Type II cells in organotypic culture can also synthesize the protein moiety of surfactant. Leucine was found to incorporate linearly into surfactant proteins for approx. 8 h.
11.
Transport as the Rate-Limiting Step in the Incorporation of Uridine into Mengovirus Ribonucleic Acid in Novikoff Rat Hepatoma Cells 总被引:3,自引:2,他引:1
The incorporation of uridine into the nucleotide pool of actinomycin-treated, mengovirus-infected Novikoff rat hepatoma cells in culture follows simple Michaelis-Menten kinetics, and the apparent V(max) and K(m) values are similar to those for uridine transport by uninfected cells. Incorporation of uridine into mengovirus-specific ribonucleic acid (RNA) also follows Michaelis-Menten kinetics, and the apparent K(m) (about 10 mum) is approximately the same as for uridine transport. Inhibition of uridine transport by the presence of adenosine, persantin, or phenethyl alcohol inhibits simultaneously and to the same extent the incorporation of uridine into the nucleotide pool and into viral RNA, without affecting viral RNA synthesis per se. Phenethyl alcohol, however, also inhibits virus maturation. The inhibition of uridine incorporation into the nucleotide pool and into viral RNA is of the simple competitive type, indicating that transport into the cells is the rate-limiting step in the incorporation of uridine into mengovirus RNA. The results also indicate that treatment with actinomycin D or mengovirus infection does not affect uridine transport. 相似文献
12.
The present study investigated the effect of insulin on phosphatidylcholine turnover in rat adipocytes labelled to equilibrium with [14C]-choline. Insulin induced a rapid turnover of this major phospholipid that was maximal by 1 min and transient in nature. Following a 1 min stimulation of the cells with insulin at a maximally effective concentration (7 nM), a 4–6% decrease in the percentage of total cellular choline associated with this phospholipid was observed. This reflected a significant transient increase in the percentage of total cellular choline associated with phosphorylcholine, which together with diacylglycerol are the phospholipase C cleavage products of phosphatidylcholine. These effects were observed over a physiological range of insulin concentrations. No effect of insulin on any other choline phospholipid or metabolite (sphingomyelin, lysophophatidylcholine, glycerophosphocholine or choline) was seen. These results suggest that insulin stimulates a phospholipase C-mediated turnover of phosphatidylcholine in rat adipocytes. The rapid nature of this turnover suggests a potential role in signal transduction. 相似文献
13.
E Yavin 《The Journal of biological chemistry》1976,251(5):1392-1397
The uptake and metabolism of [14C]choline in dissociated rat brain embryo cell cultures was examined as a function of the extracellular choline concentration. Choline uptake did not follow normal Michaelis-Menten kinetics, but rather exhibited two components with apparent Km of 0.016 mM and 0.96 mM. At low choline concentrations (high affinity uptake) most of the [14C]choline label was present in the phosphocholine fraction prior to the appearance of label in phospholipids. At high choline concentrations (low affinity uptake) a large proportion of the radioactivity was converted into acetylcholine. The dissimilarities between the formation of phosphocholine and acetylcholine as a function of choline concentration might be explained by the existence of two mutually independent enzymatic activities with different Km affinities for choline. Kinetic data augmented by double label studies, suggested that formation of choline phosphoglyceride proceeds entirely via a phosphocholine intermediate. Nearly all radioactivity in the lipid fraction is incorporated into choline phosphoglycerides. A higher turnover rate of choline incorporation into choline phosphoglycerides, accompanied by an increase in the levels of glycerophosphocholine, was observed in older cultures as compared to younger cultures. The metabolic implications of these findings in cultured brain cells in comparison with other in vitro systems are discussed. 相似文献
14.
The synthesis of phosphatidylcholine by adult rat lung alveolar type II epithelial cells in primary culture 总被引:3,自引:0,他引:3
1. The formation of phosphatidylcholine from radioactive precursors was studied in adult rat lung alveolar type II epithelial cells in primary culture. 2. The incorporation of [Me-14C]choline into total lipids and phosphatidylcholine was stimulated by addition of palmitate, whereas the incorporation of [U-14C]glucose into phosphatidylcholine and disaturated phosphatidylcholine was stimulated by addition of choline. Addition of glucose decreased the absolute rate of incorporation of [1(3)-3H]glycerol into total lipids, phosphatidylcholine and disaturated phosphatidylcholine, decreased the percentage [1(3)-3H]glycerol recovered in phosphatidylcholine, but increased the percentage phosphatidylcholine label in the disaturated species. 3. At saturating substrate concentrations, the percentages of phosphatidylcholine radioactivity found in disaturated phosphatidylcholine after incubation with [1-(14)C]acetate (in the presence of glucose) [1-(14)C]palmitate (in the presence of glucose), [Me-14C]choline (in the presence of glucose and palmitate) and [U-14C]glucose (in the presence of choline and palmitate) were 78, 75, 74 and 90%, respectively. 4. Fatty acids stimulated the incorporation of [U-14C]glucose into the glycerol moiety of phosphatidylcholine. The degree of unsaturation of the added fatty acids was reflected in the distribution of [U-14C]glucose label among the different molecular species of phosphatidylcholine. It is suggested that the glucose concentration in the blood as related to the amount of available fatty acids and their degree of unsaturation may be factors governing the synthesis of surfactant lipids. 相似文献
15.
Organotypic cultures of fetal type II epithelial cells were incubated in media containing insulin at concentrations ranging from 10 to 400 microunits/ml. Exposure to insulin resulted in increased glucose uptake from the media and in the rate of glucose conversion to CO2. Furthermore, both glucose uptake and CO2 production were dependent on the glucose concentration in the media. Surfactant and residual phosphatidylcholine fractions were isolated from the organotypic cultures by sucrose density centrifugation. The presence of low doses of insulin (10-25 microunits/ml) caused a significant increase in the incorporation of glucose into both surfactant and residual phosphatidylcholine. Insulin at levels of 100 microunits/ml or higher resulted in a significant decrease in glucose incorporation into both phosphatidylcholine fractions. Increasing the media glucose concentration from 5.6 to 20 mM caused a 2- to 2.5-fold increase in glucose utilization for surfactant and residual phospholipid synthesis, but did not produce any significant changes in choline incorporation into either surfactant or residual phosphatidylcholine. The addition of 400 microunits/ml of insulin to media containing 20 mM glucose, however, resulted in a 20% decrease in choline incorporation into surfactant phosphatidylcholine but had no effect on choline incorporation into residual phosphatidylcholine. These results suggest that insulin is an important hormone regulating fetal lung maturation and that hyperinsulinemia may be responsible for the delayed lung development in infants of diabetic mothers. 相似文献
16.
The metabolism of the phosphonium analogue of choline in cultured cells. A useful nuclear-magnetic-resonance probe for membrane phosphatidylcholine.
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1. Replacement of choline by the phosphonium analogue does not affect the growth rate of P815Y, NIL, 3T3, and SV40/3T3 cells in culture. 2. The fatty acid composition of the resulting phosphonium phosphatidylcholine is similar to that of phosphatidylcholine. 3. The rate of synthesis and degradation of phosphatidylcholine and of the phosphonium analogue are similar. 4. Phospholipid-exchange protein does not distinguish between phosphatidylcholine and the phosphonium analogue. 5. By contrast, incorporation of phosphonium choline into sphingomyelin occurs to only a minor extent. 6. It is concluded that, since the enzymes involved in the turnover of phosphatidylcholine do not discriminate between quaternary N and quaternary P in the polar head-group region, phosphonium choline should prove to be a useful probe for 31P nuclear-magnetic-resonance (n.m.r.) studies of natural membranes. 相似文献
17.
Adenine, guanine, and hypoxanthine were rapidly incorporated into the acid-soluble nucleotide pool and nucleic acids by wild type Novikoff cells. Incorporation followed normal Michaelis-Menten kinetics, but the following evidence indicates that specific transport processes precede the phosphoribosyltransferase reactions and are the rate-limiting step in purine incorporation by whole cells. Cells of an azaguanine-resistant subline of Novikoff cells which lacked hypoxanthine-guanine phosphoribosyltransferase activity and failed to incorporate guanine or hypoxanthine into the nucleotide pool, exhibited uptake of guanine and hypoxanthine by a saturable process. Similarly, wild type cells which had been preincubated in a glucose-free basal medium containing KCN and iodoacetate transported guanine and hypoxanthine normally, although a conversion of these purines to nucleotides did not occur in these cells. The mutant and KCN-iodoacetate treated wild type cells also exhibited countertransport of guanine and hypoxanthine when preloaded with various purines, uracil, and pyrimidine nucleosides. The cells also possess a saturable transport system for uracil although they lack phosphoribosyltransferase activity for uracil. In the absence of phosphoribosylation, none of the substrates was accumulated against a concentration gradient. Thus transport is by facilitated diffusion (nonconcentrative transport). Furthermore, the apparent Km values for purine uptake by untreated wild type and azaguanine-resistant cells were higher and the apparent Vmax values were lower than those for the corresponding phosphoribosyltransferases... 相似文献
18.
Fatty acid specificity for the synthesis of triacylglycerol and phosphatidylcholine and for the secretion of very-low-density lipoproteins and lysophosphatidylcholine by cultures of rat hepatocytes. 总被引:3,自引:3,他引:0
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1. The synthesis and secretion of glycerolipids by monolayer cultures of rat hepatocytes was measured by using radioactive choline, glycerol and fatty acids and by measuring the concentration of triacylglycerols in the cells. 2. The incorporation of glycerol into triacylglycerol and the accumulation of this lipid in hepatocytes showed little specificity for fatty acids, except for eicosapentaenoate, which stimulated least. Oleate was more effective at stimulating triacylglycerol secretion than were palmitate, stearate, arachidonate and eicosapentaenoate. 3. Linoleate, linolenate, arachidonate and eicosapentaenoate stimulated the incorporation of glycerol and choline into phosphatidylcholine that was secreted into the medium. By contrast, palmitate and stearate produced relatively high incorporations into the phosphatidylcholine that remained in the cells. 4. The incorporation of glycerol and choline into lysophosphatidylcholine in the medium was stimulated 2-3-fold by all of the unsaturated fatty acids tested, whereas palmitate and stearate failed to stimulate if the acids were added separately. When 1 mM-stearate was added with 1 mM-linoleate, the incorporation of linoleate into lysophosphatidylcholine was about 4 times higher than that of stearate. 5. It is proposed that the secretion of lysophosphatidylcholine by the liver could provide a transport system for choline and essential unsaturated fatty acids to other organs. 相似文献
19.
Effect of Cycocel Derivatives and Gibberellin on Choline Kinase and Choline Metabolism 总被引:2,自引:0,他引:2
Cycocel stimulated the activity of partial purified choline kinase from spinach or squash leaves, but it inhibited the activity of yeast choline kinase. The activity of different Cycocel analogs on plant growth corresponded to their stimulatory effect on the isolated choline kinase. Cycocel had no effect upon the activity of a plant phosphatase which hydrolyzed phosphorylcholine nor upon adenosine triphosphatase from wheat roots or leaves.Gibberellin A(3) inhibited choline kinase activity and reversed the stimulatory effect of Cycocel on the kinase.Total choline kinase activity per squash plant was not greatly increased by Cycocel treatment. However, on the basis of fresh weight, total kinase activity was increased by Cycocel treatment. Gibberellin A(3) partially reversed these increases. Treatment with Cycocel plus indoleacetic acid resulted in a large increase in choline kinase activity.The same distribution of tracer among phosphorylcholine, choline and betaine was observed when either phosphorylcholine-C(14) or choline-C(14) was fed to barley or wheat roots. Cycocel stimulated the incorporation of choline-C(14) into the insoluble fraction and into lipids. Cycocel inhibited phosphorylcholine uptake by roots.Thus Cycocel stimulated choline kinase activity and the utilization of choline-C(14). The effect of Cycocel upon kinase activity in vivo and in vitro was reversed by gibberellin A(3). 相似文献
20.
The coupling of choline transport to acetylcholine synthesis has been investigated by measurement of the isotopic dilution of a pulse of [3H]choline during its incorporation into the recently synthesised acetylcholine of cerebral cortex synaptosomes. Recently synthesised acetylcholine was identified as that containing 14C-labelled precursors introduced by a preincubation before the pulse. When [14C]glucose was used to label acetyl-CoA coupling ratios (calculated as the inverse of the dilution of extracellular [3H]choline during its incorporation into [3H]acetylcholine) of about 0.05-0.2 were found at a choline concentration of 1 microM, rising to 0.5 at choline concentrations of 10-50 microM. Experiments using [14C]choline as a precursor gave similar results, and it was shown that the isotopic dilution did not occur extrasynaptosomally and was not affected by low glucose concentrations. Coupling ratios were always less than unity and rose as the choline concentration increased. It is concluded that choline transported into the nerve terminal has no privileged access to choline acetyltransferase. The results can be explained by a rate-controlling transport of choline into the terminal followed by its rapid acetylation rather than any linkage or coupling of the two processes. 相似文献