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1.
L-Methionine-DL-sulfoximine (MSO), a specific inhibitor of glutaminesynthetase (GS), severely inhibited germination of New York515 Improved and of Grand Rapids lettuce seeds. However, theelongation of excised axes was not affected by MSO. MSO (1 mM)decreased the growth potential of the axes to a level nearlyequal to the restraining force of the seed coats (about 0.4M). When seeds were treated with MSO at 25?C free ammonia accumulatedin the seeds. MSO inhibited germination of several other light-sensitiveor light-hard seeds, but not light-indifferent seeds. (Received June 10, 1983; Accepted October 6, 1983)  相似文献   

2.
Experiments were conducted to investigate the effects of the convulsant L-methionine-DL-sulfoximine (MSO) on striatal dopamine (DA) metabolism. Intraventricular injections of MSO produced a transient increase in striatal DA release followed by inhibition of DA release for up to 3 days, which paralleled the inhibition by MSO of the enzyme glutamine synthetase (GS). DA synthesis was decreased for up to 24 h after injection of MSO, but returned to normal within 3 days after MSO administration. Intrastriatal injections of MSO produced a pronounced decrease in striatal DA release and inhibition of striatal GS activity 24 h postinjection but, unlike intraventricular MSO, did not produce behavioral convulsions. Glutamate-DA interactions may be responsible for the observed effects.  相似文献   

3.
Abstract: This study examines the consequences on cerebral polyamine biosynthesis of increases and decreases in cerebral methylation. Increases were elicited by administering the convulsant agent methionine sulfoximine (MSO) and decreases by elevating in vivo the cerebral levels of the methylation inhibitor S -adenosyl-homocysteine. Following the intraventricular (i.vt.) administration of one of the two possible polyamine precursors, [1,4-14C]putrescine, the specific radioactivity (sra) of the newly formed [14C]spermidine remained unchanged. Conversely, after i.vt. l -[3,4-14C]methionine, the other polyamine precursor, significantly higher sra values for [14C]spermidine and [14C]spermine were recorded in the brains of the MSO-treated animals. [14C] S - adenosylmethionine in the brain of the MSO-treated animals was also more highly labeled following [1-14C]-methionine, indicating its accelerated formation relative to controls. We also investigated the effect of the administration of adenosine + homocysteine, a treatment that results in elevated brain adenosylhomocysteine levels, on polyamine biosynthesis from [3,4-14C]-methionine. The results of these experiments show both significantly lower sra values for [14C]spermidine and [14C]spermine and significantly higher than control endogenous methionine levels, a clear sign of the existence of a retardation in the conversion of methionine to polyamines under these conditions. In conclusion, the present study demonstrates that while interference with cerebral methylation results in significant alterations of the rate of formation of the methionine moiety of spermidine and spermine, it has no effect on the entry of the putrescine moiety into the two polyamine molecules.  相似文献   

4.
Di Giacomo  C.  Sorrenti  V.  Acquaviva  R.  Campisi  A.  Vanella  G.  Perez-Polo  J. R.  Vanella  A. 《Neurochemical research》1997,22(9):1145-1150
Excessive activation of glutamate receptors via the N-methyl-D-aspartate (NMDA) subtype appears to play a role in the sequence of cellular events which lead to irreversible ischemic damage to neurons. Furthermore, NMDA receptor activation induces a stimulation of ornithine decarboxylase (ODC), the rate-limiting enzyme for polyamine (PA) biosynthesis. In order to better understand the role of PA we have measured ODC activity and the effect of methionine sulfoximine (MSO), a molecule able to stimulate ODC, on a model of transient cerebral ischemia. There was a significant increase in ODC activity in the rat cerebral cortex during post-ischemic reperfusion. The treatment with MSO induced a significant decrease in cerebral glutamine synthetase activity accompained by a marked increase in ODC activity. In MSO-pretreated rats there was a significant decrease in the survival rate when compared to untreated ischemic rats.  相似文献   

5.
At least two pathways exist in Klebsiella aerogenes for glutamate synthesis. A mutant blocked in one pathway due to the loss of glutamate dehydrogenase (gltD) does not require glutamate and has the same growth characteristics as the parent strain in most media; however, its growth is inhibited by the analogues methionine sulfoximine and methionine sulfone. Wild-type Klebsiella is resistant to 0.1 M methionine sulfoximine or methionine sulfone, whereas the gltD mutant is sensitive to 1 mM concentrations. Either glutamate or glutamine is effective in overcoming this inhibition. Activities of both glutamine synthetase and glutamate synthetase, two enzymes involved in the second pathway of glutamate synthesis, are inhibited by methionine sulfoximine and methionine sulfone. The primary effect of methionine sulfoximine appears to be the prevention of glutamine production necessary for subsequent glutamate synthesis via glutamate synthetase enzyme.  相似文献   

6.
Methionine sulfoximine (MSX), a glutamine synthetase inhibitor,suppressed the inhibitory effect of ammonia on nitrate uptakeby Anabaena cells in both the light and dark. In the light,MSX did not inhibit nitrate uptake in the absence of ammonia,but under dark conditions, MSX above 2 µM inhibited nitrateuptake. Nitrite uptake, which is not affected by ammonia ineither the light or the dark, was inhibited by MSX in the darkbut not in the light. (Received October 3, 1984; Accepted April 22, 1985)  相似文献   

7.
A number of glutamine auxotrophs of Salmonella typhimurium were isolated and characterized genetically. Three of the mutations appear to be closely linked and are complemented by episomes carrying the glnA region of Escherichia coli. The lesions in these strains are approximately 20% linked by P1 transduction with a mutation in the rha gene, but are unlinked to ilv. Another mutation causing glutamine auxotrophy in strain JB674 is genetically distinct from the others. Strain JB674 grown in glucose medium containing ammonia as the nitrogen source has reduced levels of glutamine synthetase that is more adenylylated than in the parent strain, suggesting that the enzyme can not be deadenylylated normally. The lesion causing glutamine auxotrophy in JB674 lies in the region corresponding to the glnB and glnE genes affecting glutamine synthetase modification in Klebsiella areogenes. Four Gln+ revertants of JB674 have glutamine synthetase activities 4 to 6 fold higher than normal. One mutation causing this increased enzyme synthesis has been shown by three-factor crosses with the glnA mutations to lie near or within the glnA gene.  相似文献   

8.
Methionine sulfoximine caused ammonia accumulation and photosyntheticrate inhibition in leaf discs from two C4 species, Zea maysL. cv. F. M. Cross (Hybrid) and Sorghum bicolor L. Moench cv.NC-70X, as well as one C3 plant species, Datura stramonium L.cv. stramonium. Similar results were obtained earlier with theC3 species Spinacia oleracea L. The effect occurred in the absenceof inorganic nitrogen reduction and was light dependent. Ammoniaaccumulation rates were similar in all four species examined.The results support a role for glutamine synthetase in leafammonia recycling in both C4 and C3 leaves. 1 Current address: Cetus Madison Corporation, 2208 Parview Road,Middleton, WI 53562, U.S.A. (Received November 2, 1981; Accepted April 28, 1982)  相似文献   

9.
The inactivation of glutamine synthetase by tabtoxinine-β-lactam, a phytotoxin produced by Pseudomonas syringae pv. tabaci, was shown to be irreversible. The chloroplast and cytosolic forms of the enzyme from pea leaves (Pisum sativum L.) were separated, purified, and found to be kinetically similar with Km values for glutamate of 6.7 and 4.3 millimolar and for ATP of 2.0 and 1.3 millimolar, respectively. Both forms were irreversibly inactivated by the toxin at equal rates. Using the chloroplast form, it was found that inactivation by tabtoxinine-β-lactam required ATP. Glutamate and low levels of ammonia (<2 millimolar) slowed the rate of inactivation, whereas high levels of ammonia (5, 20, and 50 millimolar) accelerated it. The inactivation proceeded at a faster rate as the pH was increased from pH 6.5 to 7.5. The role which cellular compartmentalization could play in the inactivation is discussed.  相似文献   

10.
A methionine sulfoximine (MSX) tolerant cell line of tobacco(Nicotiana tabacum L. cv. Xanthi) cells was selected by culturingthe wild-type cells in suspension media in the presence of MSXat a step-wise increase in its concentration (0.3 to 5 µM).Fifty per cent inhibition of growth occurred at 0.18 µMMSX for the wild-type cells whereas 4.65 µM was requiredfor the tolerant cells. The tolerant cells possessed about 1.5-foldincrease in glutamine synthetase (GS) activity. Kinetic experimentsshowed that an inhibitor constant for MSX was identical betweenGS isolated from these two cell types. Subunit polypeptidesof GS in both cell types were analyzed with an immunoblottingmethod by using polyclonal antibody raised against a chloroplasticGS in spinach. A single polypeptide (41 kDa) was recognizedby the antibody in wild-type cells, whereas two predominantpolypeptides of 41 and 40 kDa were seen in the MSX tolerantcells. When the GS subunit polypeptides in the wild-type cellswere examined with two-dimensional gel electrophoresis, twomajor and four minor polypeptides associating distinct chargeat 41 kDa were detected. The extract from the MSX-tolerant cellshad the same set of polypeptides at 41 kDa and in addition twomajor and some minor spots at 40 kDa. These results indicatethat 1) tobacco GS is consisted of heterogeneous subunit polypeptidesin surface charge and 2) MSX causes formation of additionalmultiple 40 kDa polypeptides which may be related to the tolerantnature of the selected cell line. (Received October 27, 1989; Accepted January 17, 1990)  相似文献   

11.
Abstract: The effect of a number of nucleotides and related compounds on glutamine synthetase (GS) induction in retina from 12-day chick embryo was studied with both biochemical and immunohistochemical techniques. A number of these compounds gave rise to GS activity comparable to that induced by treatment with cortisol, which is known to give rise to precocious induction of the enzyme in this system. Of the cyclic nucleotides examined, cAMP (0.5-1.2 × 10?3 M) gave essentially no increase in GS activity. In contrast, dibutyryl cAMP (0.8 × 10?3 M) had a more significant effect on GS activity, as did 8-bromo-cAMP and cGMP at the same concentration. The activity elicited by these nucleotides was generally half that obtained by treatment with 2.8 × 10?7 M-cortisol for the same length of time, 8-Bromo-cGMP (0.8 × 10?3 M) had an effect comparable to the aforementioned concentration of cortisol. Since phosphodiesterase activity is minimal in the chick retina at 12 days of development, addition of isobutylmethylxanthine (1 × 10?5 M) to this system had, as would be expected, little effect on GS activity. Of the noncyclic compounds, 8-bromoguanosine often gave rise to GS activity comparable to that obtained with cortisol. The other compounds (8-bromo-5′-GMP, guanosine, adenosine, and 5′-AMP) generally had less of an effect on GS. In general, the degree of staining in the immunohistochemical localization of GS corresponded well with the biochemical results and showed the enzyme to be present in regions consistent with the distribution of Muller cells and their processes. Thin-layer chromatography and radioimmunoassay of the nucleotides did not show any steroid impurity in any of the compounds used in the study, even when determinations were carried out at five times the concentration of nucleotide used in the experiments.  相似文献   

12.
Methionine sulfoximine (MSO) greatly reduced the carbon dioxideexchange rate (CER) of detached wheat (Triticum aestivvm L.cv Roland) leaves in 21% O2, but only slightly reduced it in2% O2. A supply of 50 mM NH4Cl had little effect on the CERirrespective of the O2 concentration. A simultaneous additionof glutamine and MSO protected against the inhibition of photosynthesisto a considerable extent and caused the accumulation of moreNH3 than did the addition of MSO alone. Fixation of 14CO2 in wheat leaves was inhibited by MSO treatmentin 22% O2, and there was decreased incorporation of 14G intoamino acids and sugars and increased label into acid fractions.The addition of MSO and glutamine together eliminated the effectof MSO on the photosynthetic 14CO2 fixation pattern. NH4Cl stimulatedthe synthesis of amino acids from 14CO2, especially the synthesisof serine in 22% O2. Our observations show that factors other than the uncouplingof photophosphorylation by accumulated NH3 may be responsiblefor the early stage of photosynthesis inhibition by MSO underphotorespiratory conditions. 1Present address: Department of Agricultural Chemistry, KyushuUniversity, Fukuoka 812 Japan. 2Also at U.S. Department of Agriculture, Agricultural ResearchService, Urbana, Illionois 61801, U.S.A. (Received September 13, 1983; Accepted February 2, 1984)  相似文献   

13.
Ammonia causes astrocyte swelling which is abrogated by methionine sulfoximine (MSO). Since myo-inositol is an important osmolyte, we investigated the effects of ammonia and MSO on myo-inositol flux in cultured astrocytes for periods up to 72 hours. Uptake of myo-inositol was significantly decreased by 26.7 (P < 0.05) and 39.3 (P lt; 0.006) percent after 48 hours of exposure to 5 or 10 mM ammonia, respectively. The maximum rate of uptake was 14.0 ± 0.5 nmol/hour/mg protein which was reduced to 7.45 ± 0.27 and 7.02 ± 0.57 nmoles/hour/mg protein by 5 or 10 mM ammonia, respectively. The Kms by Michaelis-Menten equation for the control, and in the presence of 5, or 10 mM ammonia were 32.5 ± 4.52, 44.4 ± 5.82, and 39.3 ± 7.0 M, respectively. Kms by Hanes-Woolf plot for the control, 5, or 10 mM ammonia were 25, 45, and 40 M, respectively. Treatment of astrocytes with either 5 or 10 mM NH4Cl for 6 hours caused a decrease in myo-inositol content by 66% and 58%, respectively. MSO (3 mM) partially diminished the ammonia-induced inhibition of myo-inositol uptake and decreased myo-inositol content by 31% after 24 hours. Additionally, ammonia increased myo-inositol efflux briefly through the fast efflux component but had little effect on myo-inositol efflux through the slow efflux component of astrocytes exposed to ammonia for up to 72 hours. Predominantly decreased myo-inositol influx coupled with brief efflux through the fast component may represent an adaptive response to diminish the extent of ammonia-induced astrocyte swelling.  相似文献   

14.
测定了水稻种子不同萌发时期胚乳、胚芽鞘和幼根的谷氨酰胺合成酶(GS)和依赖于NADH的谷氨酸合酶(NADH-GOGAT)活性变化。胚乳和胚芽鞘的GS活性在萌发过程中升高,幼根的GS活性则有所降低。NADH-GOGAT的活性变化趋势与GS相同。Native-PAGE活性染色表明,在萌发阶段的水稻种子胚乳和幼根里,始终只观察到一种GS活性带。但是,在水稻种子萌发3d后,在胚芽鞘中除继续检测到GS1的活性外,还可以观察到GS2的活性。蛋白质印迹显示,水稻种子胚乳中的GS(GSe)和GS1和GSra一样是一种胞质型GS。实验结果提示,这些不同组织中的GS与NADH-GOGAT构成的循环途径也许是水稻种子萌发时氨同化的主要途径。  相似文献   

15.
The effect of administration of chlorpromazine on the activity of glutamine synthetase and glutaminase and the content of glutamate and gamma-aminobutyric acid (GABA) in different regions of rat brain was studied in an investigation of the possible role of these amino acids in the lowering of the seizure threshold following prolonged administration of chlorpromazine. Chlorpromazine was administered at a dose of 20 mg/kg of body weight s.c. For the acute study, the animals were killed 20 min after a single injection. For the long-term study, the animals were treated every day with the same dose for 21 days and were killed 20 min after the last injection. The results showed an increase in glutamate level in each brain region investigated following long-term administration, but only in the cerebral cortex after a single dose. GABA levels showed an increase in the brainstem only in acute experiments. Glutamine synthetase activity was increased in all three regions after a single dose and only in cerebral cortex after long-term administration. Glutaminase activity showed a decrease in cerebral cortex only after long-term administration of the drug. These results suggest the possible occurrence of a state of increased excitability in the brain as a result of long-term administration of chlorpromazine, thus contributing to the known complication of seizures.  相似文献   

16.
Methionine sulfoximine inhibits the growth of Salmonella typhimurium at a concentration of 50 muM, and the addition of glutamine, but not glutamate, is sufficient to overcome this inhibition. The analogue causes 50% inhibition of glutamine synthetase activity at 2 to 4 muM and of glutamate synthase at 2 to 3 mM when these enzymes are assayed in vitro. No inhibition of glutamate dehydrogenase activity is observed at analogue concentrations as high as 50 mM. Two mutants selected for their resistance to methionine sulfoximine inhibition have a partial growth requirement for glutamine and a reduction in the glutamine synthetase and glutamate synthase activities. The sensitivity of the remaining glutamine synthetase activity in these mutants to methionine sulfoximine inhibition appears unaltered, and the lesions conferring the analogue resistance may not affect glutamine synthetase directly.  相似文献   

17.
Mutant barley plants containing only 8%, 16% or 38% of the wildtype level of glutamine synthetase activity have been isolated.The level of glutamine synthetase activity in the roots of themutant containing only 8% leaf activity was not affected bythis mutation. The plants accumulated high levels of ammoniain leaves exposed to air and although they were able to carryout photosynthetic CO2fixation normally at low levels of atmosphericO2, they were unable to maintain wild type rates of CO2fixationin air. The extent of this inhibition and the extent to whichammonia accumulated in the leaves was dependent on the photonfluence rate intercepted by the plant. When leaves from themutant plant were fed glutamine under non-photorespiratory conditionsfor 40 min before they were transferred to air, the plants exhibitedwild type rates of CO2 fixation in air but the ammonia contentof the leaves increased to an even higher level. At least inthe short term, therefore, ammonia accumulation was not responsiblefor the dramatic decline in the fixation rate of these mutantsin air. The most probable explanation is that as the supplyof potential amino donors diminished on transfer to air, therewas a restriction on the return of glycerate to the Calvin cyclewithin the chloroplast. Key words: Ammonia toxicity, photorespiration, photosynthesis, GS-deficient barley  相似文献   

18.
In soyabeans (Glycine max (L.) Merr.), glutamine synthetase(GS) activity was greater at 28 °C than at 35 °C. Thereverse was true for foliar-N loss. In field-grown plants at29 °C, the GS activity was higher and foliar-N loss waslower in soyabeans than in Amaranthus palmeri (S.) Wats. Methioninesulphoximine, a GS inhibitor, and 6-diazo-5-oxo-L-norleucine,a glutamate synthase inhibitor, significantly increased foliar-Nloss from soyabeans. The data suggest that conditions conduciveto decreased GS or glutamate synthase activity may result inincreased foliar-N loss. Palmer amaranth, Amaranthus palmeri, soyabean, Glycine max, methionine sulphoximine, 6-diazo-5-oxo-L-norleucine  相似文献   

19.
Abstract: The glutamine cycle has been proposed as a pathway in which glutamine synthesized in glia provides substrate for synthesis of the neurotransmitters glutamate and GABA as they are lost from neurons. To test whether GABA may regulate this pathway, the effect of elevated GABA on the glial enzyme glutamine synthetase was examined in rat brain. Repeated subcutaneous injections of the antiepileptic GABA transaminase inhibitor γ-vinylGABA at a dose of 150 mg/kg per day for 21 days reduced glutamine synthetase activity by 36% in the cortex and 22% in the cerebellum. At 30 mg/kg per day, glutamine synthetase activity was reduced by 9.5% in the cortex but unchanged in the cerebellum. The reductions were brain specific because the skeletal muscle and liver enzymes were unaffected by γ-vinylGABA administration. Amino acid analysis of the cortex from γ-vinylGABA-treated rats demonstrated a 270% increase in GABA levels after 150 mg/kg but no change after 30 mg/kg. GABA levels and glutamine synthetase activity were inversely correlated. The 150 mg/kg dose significantly lowered cortical glutamine and glutamate levels. The decline in brain glutamine synthetase activity with chronic γ-vinylGABA administration developed gradually over time and may be due to the slow turnover of this enzyme in vivo.  相似文献   

20.
Ethylene, used as a stimulant of latex production in Hevea brasiliensis, significantly activates the regenerating metabolism within the laticiferous cells. In this context, attention was focused on glutamine synthetase (GS; EC 6.3.1.2), a key enzyme in nitrogen metabolism. A specific and significant activation of the cytosolic glutamine synthetase (GScyt) in the laticiferous cells after ethylene treatment parallels the increase of latex yield. A marked accumulation of the corresponding mRNA was found, but in contrast, a slight and variable increase of the polypeptide level is at the limit of detection by western blotting. The GS response to ethylene might be mediated by ammonia that increases in latex cytosol following ethylene treatment. The physiological significance for such a regulation by ethylene of the GScyt is discussed in terms of the nitrogen requirement for protein synthesis associated with latex regeneration.  相似文献   

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