METABOLISM OF C14 AMINO ACIDS AND AMIDES IN DETACHED LEAVES

1956 ◽  
Vol 34 (4) ◽  
pp. 423-433 ◽  
Author(s):  
C. D. Nelson ◽  
G. Krotkov
Keyword(s):  
Amino Acids ◽  
Double Bond ◽  
Glutamic Acid ◽  
Broad Bean ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Activity Data ◽  
Acid Cycle ◽  
Bean Leaves

Detached broad bean leaves were placed with their petioles in 0.01 M ammonium nitrate and allowed to carry on photosynthesis in C14O2 for various periods from 12 to 125 min. The radioactivities of the various amino acids formed from C14O2 were determined. In addition, these amino acids were degraded by decarboxylation with ninhydrin. From the specific activity data it was concluded that the amino acid closest to the site of carbon dioxide fixation in photosynthesis was alanine, followed by aspartic and glutamic acids, with the amides farthest removed. From the intramolecular distribution of label it was concluded that asparagine and glutamine were formed from their corresponding amino acids. The labelling in aspartic and glutamic acids was not consistent with the view that these two amino acids are formed from their corresponding α-keto acids produced by operation of the conventional tricarboxylic acid cycle. A C2 plus C2 condensation is postulated for the formation of aspartic acid. A shift in the double bond in the aconitase reaction of the tricarboxylic acid cycle would account for the observed labelling in glutamic acid. When acetate-1-C14 was fed to detached broad bean leaves in the light or dark, the distribution of label in glutamic acid supported the suggestion that there is such a. shift in the double bond in the aconitase reaction. Sodium arsenite, infiltrated into tobacco leaves, inhibited the biosynthesis of asparagine but not that of glutamine.

Studies on wheat plants using 14C-labelled compounds. XXII. Incorporation into wheat protein

1969 ◽  
Vol 47 (1) ◽  
pp. 19-23 ◽  
Author(s):  
W. B. McConnell
Keyword(s):  
Glutamic Acid ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Radioactive Tracers ◽  
Wheat Protein ◽  
Acid Cycle ◽  
Late Stages ◽  
Kernel Protein

Thatcher wheat plants were labelled with 14C by injecting radioactive tracers into the top internode of the stem during late stages of plant growth. The distribution of 14C in fully mature plants was examined, emphasis being placed on the labelling of kernel-protein amino acids.Glutamine was only slightly more effective than glutamic acid for labelling glutamic acid isolated from the gluten hydrolysate, indicating that glutamic acid and glutamine are extensively interconverted in the wheat plants. Proline and glutamic acid also are readily interconverted, proline-14C yielding protein in which the glutamic acid has a higher specific activity than does the proline. By contrast, arginine-5-14C did not yield highly labelled glutamic acid.14C from glyoxylate-1-14C was widely distributed among kernel components but it produced glycine and serine with carboxyl carbons of exceptionally high specific activity.Succinate-1,4-14C, succinate-2,3-14C, and aspartic acid-14C all labelled glutamic acid of kernel protein more extensively than the other amino acids of the protein. The role of the tricarboxylic acid cycle in utilizing these tracers is discussed.

scholarly journals Metabolic Activity Related to the Potassium Pump in the Midgut of Bombyx Mori Larvae

1985 ◽  
Vol 116 (1) ◽  
pp. 69-78
Author(s):  
P. PARENTI ◽  
B. GIORDANA ◽  
V. F. SACCHI ◽  
G. M. HANOZET ◽  
A. GUERRITORE
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Glutamic Acid ◽  
Amino Acid Metabolism ◽  
Acid Metabolism ◽  
Tricarboxylic Acid Cycle ◽  
Electrical Potential ◽  
Tricarboxylic Acid ◽  
Electrical Potential Difference ◽  
Acid Cycle

The transepithelial electrical potential difference across the isolated midgut of Bombyx mori larvae is dependent on the presence of potassium and is unaffected by the addition of hexoses to perfusion media, whereas it is enhanced by alanine, aspartic acid, glutamic acid and the corresponding 2- oxoacids, glutamine and malate. The midgut enzyme profile indicates that the substrates for the tricarboxylic acid cycle are supplied mainly by amino acid metabolism via transaminases. Accordingly, aminoxyacetate drastically reduces the intestinal transepithelial electrical potential difference stimulated by amino acids. Measurement of the free amino acid concentration in the lumen content, intestinal cells and haemolymph shows that glutamic acid, asparagine and glutamine are accumulated in the cell, whilst the haemolymph is enriched with basic amino acids and with glycine, alanine, serine and tyrosine, the major components of the silk fibroin. Therefore, amino acid metabolism directly related to the tricarboxylic acid cycle seems to be the primary source of energy for the potassium pump activity in B. mori midgut.

STUDIES ON WHEAT PLANTS USING C14 COMPOUNDS: V. GERMINATION STUDIES WITH LABELLED WHEAT SEEDS

1957 ◽  
Vol 35 (1) ◽  
pp. 1259-1266 ◽  
Author(s):  
W. B. McConnell
Keyword(s):  
Carbon Dioxide ◽  
Amino Acid ◽  
Glutamic Acid ◽  
Active Compound ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
Carbon 14 ◽  
Wheat Seeds

Radioactive wheat seeds, obtained by injecting acetate-C14 into the stems of the parent plants, were germinated in the absence of light and nutrient and the fate of the carbon-14 was observed. Carbon respired as carbon dioxide had a higher specific activity than any of the major seed components except protein. Variations were found in the patterns by which material was transferred from the kernel to new tissue as reflected in a comparison of the activity of various components. Glutamic acid was the most active compound isolated either from the original seeds or from the new tissues. This observation, together with similarities noted in the intramolecular distribution of carbon-14 in glutamic acid of new tissue and seed residues, indicated that glutamic acid was reutilized for the biosynthesis of seedling protein. Changes in the labelling of glutamic acid during transfer to new tissue are qualitatively in accord with the idea that at least some of the amino acid is used after re-entry into the tricarboxylic acid cycle.

STUDIES ON WHEAT PLANTS USING C14 COMPOUNDS: V. GERMINATION STUDIES WITH LABELLED WHEAT SEEDS

1957 ◽  
Vol 35 (12) ◽  
pp. 1259-1266 ◽  
Author(s):  
W. B. McConnell
Keyword(s):  
Carbon Dioxide ◽  
Amino Acid ◽  
Glutamic Acid ◽  
Active Compound ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
Carbon 14 ◽  
Wheat Seeds

Radioactive wheat seeds, obtained by injecting acetate-C14 into the stems of the parent plants, were germinated in the absence of light and nutrient and the fate of the carbon-14 was observed. Carbon respired as carbon dioxide had a higher specific activity than any of the major seed components except protein. Variations were found in the patterns by which material was transferred from the kernel to new tissue as reflected in a comparison of the activity of various components. Glutamic acid was the most active compound isolated either from the original seeds or from the new tissues. This observation, together with similarities noted in the intramolecular distribution of carbon-14 in glutamic acid of new tissue and seed residues, indicated that glutamic acid was reutilized for the biosynthesis of seedling protein. Changes in the labelling of glutamic acid during transfer to new tissue are qualitatively in accord with the idea that at least some of the amino acid is used after re-entry into the tricarboxylic acid cycle.

The aerobic utilization of pyruvate in plant tissues

1959 ◽  
Vol 150 (939) ◽  
pp. 192-198 ◽  
Keyword(s):  
Amino Acids ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Methyl Groups ◽  
Plant Tissues ◽  
Root Tips ◽  
Acid Cycle ◽  
Amino Butyric Acid ◽  
Apple Tissue

Pyruvate with 14 C-labelling in the carbonyl and methyl groups was supplied to apple tissue and to root tips of barley. After incubation the labelling was found in a series of carboxylic acids and in alanine, glutamic acid, aspartic acid and γ -amino-butyric acid. Glutamine and asparagine were also labelled; but several other amino acids whose presence was demonstrated were without label after 4 h. Sugars and polysaccharides were also unlabelled. The CO 2 given off invariably contained 14 C, but the specific activity was much lower than that of the pyruvate supplied. It is concluded that the fed pyruvate only very partially replaced internal substrates and that it was oxidized in a tricarboxylic acid cycle. It gave rise to alanine by direct amination and to other amino acids after partial oxidation. No pyruvate was built back to sugars or other carbohydrates in either tissue.

scholarly journals Utilization in vivo of glucose and volatile fatty acids by sheep brain for the synthesis of acidic amino acids

1966 ◽  
Vol 101 (3) ◽  
pp. 591-597 ◽  
Author(s):  
R M O'Neal ◽  
R E Koeppe ◽  
E I Williams
Keyword(s):  
Amino Acids ◽  
Fatty Acids ◽  
Glutamic Acid ◽  
Aspartic Acid ◽  
Free Amino Acids ◽  
Free Amino ◽  
Specific Activity ◽  
Tricarboxylic Acid Cycle ◽  
Sheep Brain ◽  
The Brain

1. Free glutamic acid, aspartic acid, glutamic acid from glutamine and, in some instances, the glutamic acid from glutathione and the aspartic acid from N-acetyl-aspartic acid were isolated from the brains of sheep and assayed for radioactivity after intravenous injection of [2-(14)C]glucose, [1-(14)C]acetate, [1-(14)C]butyrate or [2-(14)C]propionate. These brain components were also isolated and analysed from rats that had been given [2-(14)C]propionate. The results indicate that, as in rat brain, glucose is by far the best precursor of the free amino acids of sheep brain. 2. Degradation of the glutamate of brain yielded labelling patterns consistent with the proposal that the major route of pyruvate metabolism in brain is via acetyl-CoA, and that the short-chain fatty acids enter the brain without prior metabolism by other tissue and are metabolized in brain via the tricarboxylic acid cycle. 3. When labelled glucose was used as a precursor, glutamate always had a higher specific activity than glutamine; when labelled fatty acids were used, the reverse was true. These findings add support and complexity to the concept of the metabolic; compartmentation' of the free amino acids of brain. 4. The results from experiments with labelled propionate strongly suggest that brain metabolizes propionate via succinate and that this metabolic route may be a limited but important source of dicarboxylic acids in the brain.

scholarly journals Tracer studies on the biosynthesis of amino acids from lactate by Peptostreptococcus elsdenii

1967 ◽  
Vol 105 (1) ◽  
pp. 299-310 ◽  
Author(s):  
H. J. Somerville ◽  
J. L. Peel
Keyword(s):  
Amino Acids ◽  
Glutamic Acid ◽  
Aspartic Acid ◽  
Tricarboxylic Acid Cycle ◽  
Carbon Chain ◽  
Tricarboxylic Acid ◽  
Diaminopimelic Acid ◽  
Reaction Sequence ◽  
Tracer Studies ◽  
Cell Fractions

Peptostreptococcus elsdenii, a strict anaerobe from the rumen, was grown on a medium containing yeast extract and [1−14C]- or [2−14C]-lactate. Radioisotope from lactate was found in all cell fractions, but mainly in the protein. The label in the protein fraction was largely confined to a few amino acids: alanine, serine, aspartic acid, glutamic acid and diaminopimelic acid. The alanine, serine, aspartic acid and glutamic acid were separated, purified and degraded to establish the distribution of 14C from lactate within the amino acid molecules. The labelling patterns in alanine and serine suggested their formation from lactate without cleavage of the carbon chain. The pattern in aspartic acid suggested formation by condensation of a C3 unit derived directly from lactate with a C1 unit, probably carbon dioxide. The distribution in glutamic acid was consistent with two possible pathways of formation: (a) by the reactions of the tricarboxylic acid cycle leading from oxaloacetate to 2-oxoglutarate, followed by transamination; (b) by a pathway involving the reaction sequence 2 acetyl-CoA→crotonyl-CoA→glutaconate→glutamate.

scholarly journals Neuronal-glial metabolism under depolarizing conditions. A 13C-n.m.r. study

1992 ◽  
Vol 282 (1) ◽  
pp. 225-230 ◽  
Author(s):  
R S Badar-Goffer ◽  
O Ben-Yoseph ◽  
H S Bachelard ◽  
P G Morris
Keyword(s):  
Amino Acids ◽  
Glial Cells ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acid Cycle ◽  
Maximum Percentage ◽  
Theoretical Maximum ◽  
Metabolic Pool ◽  
Time Courses ◽  
The Individual

Time courses of incorporation of 13C from 13C-labelled glucose and/or acetate into the individual carbon atoms of amino acids, citrate and lactate in depolarized cerebral tissues were monitored by using 13C-n.m.r. spectroscopy. There was no change in the maximum percentage of 13C enrichments of the amino acids on depolarization, but the maxima were reached more rapidly, indicating that rates of metabolism in both glycolysis and the tricarboxylic acid cycle were accelerated. Although labelling of lactate and of citrate approached the theoretical maximum of 50%, labelling of the amino acids was always below 20%, suggesting that there is a metabolic pool or compartment that is inaccessible to exogenous substrates. Under resting conditions labelling of citrate and of glutamine from [1-13C]glucose was not detected, whereas both were labelled from [2-13C]acetate, which is considered to reflect glial metabolism. In contrast, considerable labelling of these two metabolites from [1-13C]glucose was observed in depolarized tissues, suggesting that the increased metabolism may be due to increased consumption of glucose by glial cells. The labelling patterns on depolarization from [1-13C]glucose alone and from both precursors [( 1-13C]glucose plus [2-13C]acetate) were similar, which also indicates that the changes are due to increased consumption of glucose rather than acetate.

scholarly journals Effect of renal tubule-specific knockdown of the Na+/H+exchanger NHE3 in Akita diabetic mice

2019 ◽  
Vol 317 (2) ◽  
pp. F419-F434 ◽  
Author(s):  
Akira Onishi ◽  
Yiling Fu ◽  
Manjula Darshi ◽  
Maria Crespo-Masip ◽  
Winnie Huang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Phosphoenolpyruvate Carboxykinase ◽  
Tricarboxylic Acid Cycle ◽  
Branched Chain Amino Acids ◽  
Tricarboxylic Acid ◽  
Proximal Tubules ◽  
Wild Type ◽  
Kidney Weight ◽  
Acid Cycle ◽  
Branched Chain

Na+/H+exchanger isoform 3 (NHE3) contributes to Na+/bicarbonate reabsorption and ammonium secretion in early proximal tubules. To determine its role in the diabetic kidney, type 1 diabetic Akita mice with tubular NHE3 knockdown [Pax8-Cre; NHE3-knockout (KO) mice] were generated. NHE3-KO mice had higher urine pH, more bicarbonaturia, and compensating increases in renal mRNA expression for genes associated with generation of ammonium, bicarbonate, and glucose (phosphoenolpyruvate carboxykinase) in proximal tubules and H+and ammonia secretion and glycolysis in distal tubules. This left blood pH and bicarbonate unaffected in nondiabetic and diabetic NHE3-KO versus wild-type mice but was associated with renal upregulation of proinflammatory markers. Higher renal phosphoenolpyruvate carboxykinase expression in NHE3-KO mice was associated with lower Na+-glucose cotransporter (SGLT)2 and higher SGLT1 expression, indicating a downward tubular shift in Na+and glucose reabsorption. NHE3-KO was associated with lesser kidney weight and glomerular filtration rate (GFR) independent of diabetes and prevented diabetes-associated albuminuria. NHE3-KO, however, did not attenuate hyperglycemia or prevent diabetes from increasing kidney weight and GFR. Higher renal gluconeogenesis may explain similar hyperglycemia despite lower SGLT2 expression and higher glucosuria in diabetic NHE3-KO versus wild-type mice; stronger SGLT1 engagement could have affected kidney weight and GFR responses. Chronic kidney disease in humans is associated with reduced urinary excretion of metabolites of branched-chain amino acids and the tricarboxylic acid cycle, a pattern mimicked in diabetic wild-type mice. This pattern was reversed in nondiabetic NHE3-KO mice, possibly reflecting branched-chain amino acids use for ammoniagenesis and tricarboxylic acid cycle upregulation to support formation of ammonia, bicarbonate, and glucose in proximal tubule. NHE3-KO, however, did not prevent the diabetes-induced urinary downregulation in these metabolites.

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