scholarly journals Effect of glucose deprivation on rat glutamine synthetase in cultured astrocytes

1996 ◽  
Vol 315 (2) ◽  
pp. 607-612 ◽  
Author(s):  
Françoise ROSIER ◽  
Dominique LAMBERT ◽  
Jeannine MERTENS-STRIJTHAGEN
Keyword(s):  
Glutamine Synthetase ◽  
Specific Activity ◽  
Glucose Deprivation ◽  
Synthesis Rate ◽  
Synthetase Activity ◽  
Glutamine Synthetase Activity ◽  
Sprague Dawley ◽  
Experimental Conditions ◽  
Adult Rats ◽  
Total Recovery

Glutamine synthetase was purified from the cerebral cortex of adult rats and characterized. Polyclonal rabbit antibodies were raised against the enzyme, purified and their specific anti-(glutamine synthetase) activity determined. A primary astroglial culture was prepared from newborn Sprague–Dawley rats. Astrocytes at different ages of development were incubated in the presence and absence of glucose. In glucose-deprived conditions the specific activity of glutamine synthetase decreased. This decrease was more pronounced in 8-day-old than in 21-day-old cultures. Kinetic analysis demonstrated that the reduction in activity was mainly related to a decrease in Vmax. By immunoprecipitation, it was shown that the number of enzyme molecules in astrocytes was decreased in glucose-deprived conditions. On addition of glucose, a total recovery of glutamine synthetase was obtained after 36 h in 8-day-old culture. Rates of degradation and synthesis were investigated. When compared with an incubation in the presence of glucose, glucose deprivation increased enzyme turnover, as estimated from the first-order disappearance of radioactivity from glutamine synthetase. Synthesis rate was estimated from the incorporation of [35S]methionine during a 2 h incubation period and was decreased in glucose-deprived conditions. Trichloroacetate-precipitable proteins changed only slightly in the experimental conditions, and total protein did not vary significantly during the experimental period. A mathematical model is presented which attempts to integrate degradation and synthesis in our experimental model.

Glutamine synthetase from Mycobacterium avium

1984 ◽  
Vol 30 (3) ◽  
pp. 353-359 ◽  
Author(s):  
Maria E. Alvarez ◽  
C. M. McCarthy
Keyword(s):  
Enzyme Activity ◽  
Glutamine Synthetase ◽  
Nitrogen Source ◽  
Mycobacterium Avium ◽  
Specific Activity ◽  
Sedimentation Coefficient ◽  
Ammonia Assimilation ◽  
Methionine Sulfoximine ◽  
Synthetase Activity ◽  
Glutamine Synthetase Activity

Mycobacterium avium was previously shown to be dependent upon ammonia or glutamine as a nitrogen source. In an effort to assess the physiology of ammonia assimilation by M. avium, a characterization of its glutamine synthetase was performed. The enzyme from M. avium was purified by streptomycin sulfate treatment, ammonium sulfate precipitation, and affinity chromatography. The enzyme was unusual in that it had a pH optimum of 6.4 and maximum enzyme activity was obtained between 50 and 60 °C as shown by the transferase assay. The glutamine synthetase activity from batch-cultured cells decreased with increasing concentration of ammonium chloride in the range of 0.25–5 μ mol/mL of medium, which demonstrated a response to environmental supply of a nitrogen source. The mycobacterial enzyme was similar to the other bacterial glutamine synthetases in terms of molecular weight and sedimentation coefficient which were 600 000 and 19.5 S, respectively, and enzyme activity was lost by treatment with a glutamate analog, methionine sulfoximine. The isoelectric point was, however, pH 4.5. Treatment of the enzyme with snake venom phosphodiesterase resulted in an increase in specific activity. AMP was released by the phosphodiesterase treatment, thus demonstrating that M. avium glutamine synthetase was regulated by adenylylation modification.

scholarly journals Chronic Valproic Acid Administration Increases Plasma, Liver, and Brain Ammonia Concentration and Suppresses Glutamine Synthetase Activity

2020 ◽  
Vol 10 (10) ◽  
pp. 759
Author(s):  
Abdelnaser A. Badawy ◽  
Rasha Elghaba ◽  
Mohamed Soliman ◽  
Abdelaziz M. Hussein ◽  
Sana A. AlSadrah ◽  
...  
Keyword(s):  
Valproic Acid ◽  
Glutamine Synthetase ◽  
Ammonia Concentration ◽  
Synthetase Activity ◽  
Glutamine Synthetase Activity ◽  
Sprague Dawley ◽  
Seizure Disorders ◽  
Sprague Dawley Rats ◽  
Underlying Mechanisms ◽  
The Brain

Asymptomatic valproic acid (VPA)-induced hyperammonemia in the absence of liver impairment is fairly common. However, the underlying mechanisms through which VPA causes elevation in plasma ammonia (NH4) remains under investigation. Male Sprague Dawley rats (n = 72) were randomly allocated to receive VPA 400 mg/kg, 200 mg/kg, or vehicle IP daily for either 8, 14, or 28 consecutive days. The behavioral effects of VPA were assessed. Plasma, liver, and prefrontal cortex (PFC), striatum (Str), and cerebellum (Cere) were collected 1 h post last injection and assayed for NH4 concentration and glutamine synthetase (GS) enzyme activity. Chronic VPA treatment caused attenuation of measured behavioral reflexes (p < 0.0001) and increase in plasma NH4 concentration (p < 0.0001). The liver and brain also showed significant increase in tissue NH4 concentrations (p < 0.0001 each) associated with significant reduction in GS activity (p < 0.0001 and p = 0.0003, respectively). Higher tissue NH4 concentrations correlated with reduced GS activity in the liver (r = −0.447, p = 0.0007) but not in the brain (r = −0.058, p = 0.4). Within the brain, even though NH4 concentrations increased in the PFC (p = 0.001), Str (p < 0.0001), and Cere (p = 0.01), GS activity was reduced only in the PFC (p < 0.001) and not in Str (p = 0.2) or Cere (p = 0.1). These results suggest that VPA-induced elevation in plasma NH4 concentration could be related, at least in part, to the suppression of GS activity in liver and brain tissues. However, even though GS is the primary mechanism in brain NH4 clearance, the suppression of brain GS does not seem to be the main factor in explaining the elevation in brain NH4 concentration. Further research is urgently needed to investigate brain NH4 dynamics under chronic VPA treatment and whether VPA clinical efficacy in treating seizure disorders and bipolar mania is impacted by its effect on GS activity or other NH4 metabolizing enzymes.

Respective effects of glucose and glutamine on the glutamine synthetase activity of human skin fibroblasts

1991 ◽  
Vol 102 (2) ◽  
Author(s):  
Th�ophile Soni ◽  
Claire Wolfrom ◽  
Samia Guerroui ◽  
Nicole Raynaud ◽  
Jos�phine Poggi ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Human Skin ◽  
Skin Fibroblasts ◽  
Synthetase Activity ◽  
Glutamine Synthetase Activity ◽  
Human Skin Fibroblasts

Control of nitrogen assimilation in Stichococcus bacillaris by growth conditions

1987 ◽  
Vol 65 (3) ◽  
pp. 432-437 ◽  
Author(s):  
Iftikhar Ahmad ◽  
Johan A. Hellebust
Keyword(s):  
Glutamine Synthetase ◽  
Nitrogen Assimilation ◽  
Continuous Light ◽  
Synthetase Activity ◽  
Glutamine Synthetase Activity ◽  
Dark Cycle ◽  
Continuous Darkness ◽  
Cell Carbon ◽  
Stichococcus Bacillaris ◽  
Mixotrophic Conditions

Stichococcus bacillaris Naeg. (Chlorophyceae) grown on a 12 h light: 12 h dark cycle divides synchronously under photoautotrophic conditions and essentially nonsynchronously under mixotrophic conditions. Photoassimilation of carbon under photoautotrophic conditions was followed by a decline in cell carbon content during the dark period, whereas under mixotrophic conditions cell carbon increased throughout the light–dark cycle. The rates of nitrogen assimilation by cultures grown on either nitrate or ammonium declined sharply during the dark, and these declines were most pronounced under photoautotrophic conditions. Photoautotrophic cells synthesized glutamine synthetase and NADPH – glutamate dehydrogenase (GDH) exclusively in the light, whereas in mixotrophic cells about 20% of the total synthesis of these enzymes during one light–dark cycle occurred in the dark. NADH–GDH was synthesized almost continuously over the entire light–dark cycle. In the dark, both under photoautotrophic and mixotrophic conditions, the alga contained more than 50% of glutamine synthetase in an inactive form, which was reactivated in vitro in the presence of mercaptoethanol and in vivo after returning the cultures to the light. The thermal stability of glutamine synthetase activity was less in light-harvested cells than in dark-harvested cells. The inactivation of glutamine synthetase did not occur in cultures growing either heterotrophically in continuous darkness or photoautotrophically in continuous light. This enzyme appears to be under thiol control only in cells grown under alternating light–dark conditions, irrespective of whether this light regime results in synchronous cell division or not.

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