scholarly journals Utilization of [15N]glutamate by cultured astrocytes

1986 ◽  
Vol 234 (1) ◽  
pp. 185-192 ◽  
Author(s):  
M Yudkoff ◽  
I Nissim ◽  
K Hummeler ◽  
M Medow ◽  
D Pleasure
Keyword(s):  
Reductive Amination ◽  
Adenine Nucleotides ◽  
Ammonia Concentration ◽  
Gas Chromatography Mass Spectrometry ◽  
Purine Nucleotide ◽  
Cultured Astrocytes ◽  
Dehydrogenase Reaction ◽  
Purine Nucleotide Cycle ◽  
Almost All ◽  
Glutaminase Activity

The metabolism of 0.25 mM-[15N]glutamic acid in cultured astrocytes was studied with gas chromatography-mass spectrometry. Almost all 15N was found as [2-15N]glutamine, [2-15N]glutamine, [5-15N]glutamine and [15N]alanine after 210 min of incubation. Some incorporation of 15N into aspartate and the 6-amino position of the adenine nucleotides also was observed, the latter reflecting activity of the purine nucleotide cycle. After the addition of [15N]glutamate the ammonia concentration in the medium declined, but the intracellular ATP concentration was unchanged despite concomitant ATP consumption in the glutamine synthetase reaction. Some potential sources of glutamate nitrogen were identified by incubating the astrocytes for 24 h with [5-15N]glutamine, [2-15N]glutamine or [15N]alanine. Significant labelling of glutamate was noted with addition of glutamine labelled on either the amino or the amide moiety, reflecting both glutaminase activity and reductive amination of 2-oxoglutarate in the glutamate dehydrogenase reaction. Alanine nitrogen also is an important source of glutamate nitrogen in this system.

scholarly journals [15N]aspartate metabolism in cultured astrocytes. Studies with gas chromatography-mass spectrometry

1987 ◽  
Vol 241 (1) ◽  
pp. 193-201 ◽  
Author(s):  
M Yudkoff ◽  
I Nissim ◽  
D Pleasure
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
De Novo ◽  
Adenine Nucleotides ◽  
Gas Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry ◽  
Cultured Astrocytes ◽  
Purine Nucleotide Cycle ◽  
Citrulline Synthesis ◽  
15N Urea

The metabolism of 2.5 mM-[15N]aspartate in cultured astrocytes was studied with gas chromatography-mass spectrometry. Three primary metabolic pathways of aspartate nitrogen disposition were identified: transamination with 2-oxoglutarate to form [15N]glutamate, the nitrogen of which subsequently was transferred to glutamine, alanine, serine and ornithine; condensation with IMP in the first step of the purine nucleotide cycle, the aspartate nitrogen appearing as [6-amino-15N]adenine nucleotides; condensation with citrulline to form argininosuccinate, which is cleaved to yield [15N]arginine. Of these three pathways, the formation of arginine was quantitatively the most important, and net nitrogen flux to arginine was greater than flux to other amino acids, including glutamine. Notwithstanding the large amount of [15N]arginine produced, essentially no [15N]urea was measured. Addition of NaH13CO3 to the astrocyte culture medium was associated with the formation of [13C]citrulline, thus confirming that these cells are capable of citrulline synthesis de novo. When astrocytes were incubated with a lower (0.05 mM) concentration of [15N]aspartate, most 15N was recovered in alanine, glutamine and arginine. Formation of [6-amino-15N]adenine nucleotides was diminished markedly compared with results obtained in the presence of 2.5 mM-[15N]aspartate.

scholarly journals Sources of ammonia for mammalian urea synthesis

1978 ◽  
Vol 176 (3) ◽  
pp. 733-737 ◽  
Author(s):  
H A Krebs ◽  
R Hems ◽  
P Lund ◽  
D Halliday ◽  
W W Read
Keyword(s):  
Glutamate Dehydrogenase ◽  
Initial Rate ◽  
Adenine Nucleotides ◽  
Rat Hepatocytes ◽  
The Other ◽  
Amino Group ◽  
Urea Synthesis ◽  
Purine Nucleotide ◽  
Carbamoyl Phosphate ◽  
Purine Nucleotide Cycle

The initial rate of incorporation of [15N]alanine into the 6-amino group of the adenine nucleotides in rat hepatocytes was about one-eighteenth of the rate of incorporation into urea. Thus the purine nucleotide cycle cannot provide most of the ammonia needed in urea synthesis for the carbamoyl phosphate synthase reaction (EC 2.7.2.5). On the other hand, contrary to the view expressed by McGivan & Chappell [(1975) FEBS Lett. 52, 1–7], the experiments support the view that hepatic glutamate dehydrogenase can supply the required ammonia.

scholarly journals The purine nucleotide cycle and ammonia formation from glutamine by rat kidney slices

1983 ◽  
Vol 212 (3) ◽  
pp. 705-711 ◽  
Author(s):  
T Strzelecki ◽  
J Rogulski ◽  
S Angielski
Keyword(s):  
Adenine Nucleotides ◽  
Rat Kidney ◽  
Glucose Production ◽  
Amino Group ◽  
Purine Nucleotide ◽  
Normal Kidney ◽  
Kidney Slices ◽  
Total Ammonia ◽  
Purine Nucleotide Cycle ◽  
Ammonia Formation

To test the significance of the purine nucleotide cycle in renal ammoniagenesis, studies were conducted with rat kidney cortical slices using glutamate or glutamine labelled in the alpha-amino group with 15N. Glucose production by normal kidney slices with 2 mM-glutamine was equal to that with 3 mM-glutamate. With L-[15N]glutamate as sole substrate, one-third of the total ammonia produced by kidney slices was labelled, indicating significant deamination of glutamate or other amino acids from the cellular pool. Ammonia produced from the amino group of L-[alpha-15N]glutamine was 4-fold higher than from glutamate at similar glucose production rates. Glucose and ammonia formation from glutamine by kidney slices obtained from rats with chronic metabolic acidosis was found to be 70% higher than by normal kidney slices. The contribution of the amino group of glutamine to total ammonia production was similar in both types of kidneys. No 15N was found in the amino group of adenine nucleotides after incubation of kidney slices from normal or chronically acidotic rats with labelled glutamine. Addition of Pi, a strong inhibitor of AMP deaminase, had no effect on ammonia formation from glutamine. Likewise, fructose, which may induce a decrease in endogenous Pi, had no effect on ammonia formation. The data obtained suggest that the contribution of the purine nucleotide cycle to ammonia formation from glutamine in rat renal tissue is insignificant.

scholarly journals The Purine Nucleotide Cycle

1972 ◽  
Vol 247 (1) ◽  
pp. 162-169 ◽  
Author(s):  
Keith Tornheim ◽  
John M. Lowenstein
Keyword(s):  
Purine Nucleotide ◽  
Purine Nucleotide Cycle

scholarly journals Heterogeneous Photo-Fenton Catalytic Degradation of Practical Pharmaceutical Wastewater by Modified Attapulgite Supported Multi-Metal Oxides

Water ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 156
Author(s):  
Manjing Lu ◽  
Jiaqi Wang ◽  
Yuzhong Wang ◽  
Zhengguang He
Keyword(s):  
Photocatalytic Oxidation ◽  
Removal Rate ◽  
Catalytic Degradation ◽  
Gas Chromatography Mass Spectrometry ◽  
Pharmaceutical Wastewater ◽  
High Concentration ◽  
Modified Attapulgite ◽  
Cod Removal Rate ◽  
Pre Treatment ◽  
Almost All

Chemical synthetic pharmaceutical wastewater has characteristics of high concentration, high toxicity and poor biodegradability, so it is difficult to directly biodegrade. We used acid modified attapulgite (ATP) supported Fe-Mn-Cu polymetallic oxide as catalyst for multi-phase Fenton-like ultraviolet photocatalytic oxidation (photo-Fenton) treatment with actual chemical synthetic pharmaceutical wastewater as the treatment object. The results showed that at the initial pH of 2.0, light distance of 20 cm, and catalyst dosage and hydrogen peroxide concentration of 10.0 g/L and 0.5 mol/L respectively, the COD removal rate of wastewater reached 65% and BOD5/COD increased to 0.387 when the reaction lasted for 180 min. The results of gas chromatography-mass spectrometry (GC-MS) indicated that Fenton-like reaction with Fe-Mn-Cu@ATP had good catalytic potential and significant synergistic effect, and could remove almost all heterocycle compounds well. 3D-EEM (3D electron microscope) fluorescence spectra showed that the fluorescence intensity decreased significantly during catalytic degradation, and the UV humus-like and fulvic acid were effectively removed. The degradation efficiency of the nanocomposite only decreased by 5.8% after repeated use for 6 cycles. It seems appropriate to use this process as a pre-treatment for actual pharmaceutical wastewater to facilitate further biological treatment.

scholarly journals The role of glutamine oxidation and the purine nucleotide cycle for adaptation of tumour energetics to the transition from the anaerobic to the aerobic state

1988 ◽  
Vol 252 (2) ◽  
pp. 381-386 ◽  
Author(s):  
Z Kovacević ◽  
D Jerance ◽  
O Brkljac
Keyword(s):  
Adenine Nucleotide ◽  
The Other ◽  
Purine Nucleotide ◽  
Purine Nucleotide Cycle ◽  
Adenine Nucleotide Pool

It is proposed that the purine nucleotide cycle and glutamine oxidation play a key role in the adaptation of tumour energetics to the transition from the anaerobic to the aerobic state. In support of this proposal, it was found that glutamine and inosine markedly increase total adenylates in the presence of oxygen, whereas the addition of hadacidin abolishes this effect. Transition of the cells from the anaerobic to the aerobic state, and vice versa, in the presence of glutamine plus inosine revealed that there are two components of the adenine nucleotide pool, one which is stable and the other which is variable and responds to the aerobic-anaerobic transition. This part of the pool undergoes degradation or resynthesis owing to activation of the enzymes of the purine nucleotide cycle. Resynthesis of the pool is accompanied by substantial net utilization of aspartate, which is produced by glutamine oxidation. This is supported by the experiments in which the cells were alternately incubated with nitrogen or oxygen, demonstrating that hadacidin significantly decreased utilization of aspartate and regeneration of ATP owing to inhibition of adenylosuccinate synthase.

scholarly journals Evidence for the presence of oligophosphoglyceroyl-ATP in rat offney

1986 ◽  
Vol 240 (2) ◽  
pp. 597-599 ◽  
Author(s):  
W L Hutchinson ◽  
P J Ratcliffe ◽  
J Mowbray
Keyword(s):  
Trichloroacetic Acid ◽  
Adenine Nucleotides ◽  
Purine Nucleotide ◽  
Purine Nucleotides ◽  
Nucleotide Content ◽  
Rat Hearts ◽  
Perfused Rat Hearts ◽  
Rapid Equilibrium

The inability to account for large systematic variations in total purine nucleotide content of perfused rat hearts led to the demonstration that the soluble adenine nucleotides are in rapid equilibrium with a highly phosphorylated hetero-oligomeric derivative whose structure appears to be 3-phospho[glyceroyl-gamma-triphospho-5′-adenosine-3′-3-phosp ho]4glyceroyl- gamma-triphospho-5′-adenosine [Hutchinson, Morris & Mowbray (1986) Biochem. J. 234, 623-627]. Analogous techniques to those used with hearts for specifically labelling tissue purine nucleotides followed by extration and purification of nucleotides from the trichloroacetic acid-precipitable fraction show the existence of a corresponding rapid equilibrium between ATP and an oligomeric tetraphosphoadenosine derivative in perfused kidneys.

Binding of adenylosuccinate synthetase to contractile proteins of muscle

1989 ◽  
Vol 257 (1) ◽  
pp. C29-C35 ◽  
Author(s):  
J. P. Manfredi ◽  
R. Marquetant ◽  
A. D. Magid ◽  
E. W. Holmes
Keyword(s):  
Ionic Strength ◽  
Dissociation Constant ◽  
Thin Filament ◽  
Contractile Proteins ◽  
Purine Nucleotide ◽  
Apparent Dissociation Constant ◽  
Thin Filaments ◽  
Adenylosuccinate Synthetase ◽  
Purine Nucleotide Cycle ◽  
Low Ionic Strength

The muscle isozyme of adenylosuccinate synthetase (AdSS), an enzyme of the purine nucleotide cycle, has previously been shown to bind to purified F-actin in buffers of low ionic strength and pH (Ogawa et al. Eur. J. Biochem. 85: 331-338, 1978). We have extended these observations by measuring the association of both crude and purified AdSS with the contractile proteins of muscle in buffers of physiological ionic strength and pH. Under these conditions, the enzyme binds to F-actin, actin-tropomyosin complexes, reconstructed thin filaments, and myofibrils but not to myosin. The apparent dissociation constant of 1.2 microM and binding maximum of 2.6 nmol enzyme/mg myofibrils indicate that binding of AdSS to myofibrils can be physiologically significant. The results suggest that AdSS in muscle may be associated with the thin filament of myofibrils.

Muscle Purine Nucleotide Cycle Enzymes in Exercise Intolerance

1998 ◽  
pp. 205-209 ◽  
Author(s):  
Maria-Grazia Operti ◽  
M.-Françoise Vincent ◽  
Jean-Marie Brucher ◽  
Georges Van den Berghe
Keyword(s):  
Exercise Intolerance ◽  
Purine Nucleotide ◽  
Purine Nucleotide Cycle
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