Nitrogen metabolism of Picea glauca. V. Metabolism of uniformly labeled 14C-L-proline and 14C-L-glutamine by dormant buds in late fall

1973 ◽  
Vol 51 (2) ◽  
pp. 359-369 ◽  
Author(s):  
D. J. Durzan

In early fall, the high levels of free arginine nitrogen in spruce buds were eventually replaced by proline nitrogen, and in late spring, glutamine nitrogen accumulated. In late October when levels of free proline nitrogen were high, bud primordia from terminal shoots were excised and exposed to uniformly labeled 14C-L-proline and 14C-L-glutamine. The main early products from 14C-L-proline were Δ1-pyrroline-5-carboxylic acid, glutamic-γ-semialdehyde, and glutamic acid. Later products included glutamine, γ-aminobutyric acid, and to a much lesser extent pyrrolidone carboxylic acid, ornithine, and arginine. In protein, radioactivity was recovered from proline, glutamic acid, and hydroxyproline.Products from 14C-glutamine were mainly glutamic and α-ketoglutaric acid as well as proline, γ-aminobutyric acid, alanine, and pyrrolidone carboxylic acid. In protein, glutamic acid, aspartic acid, and proline contained carbon-14. Results indicated that proline and glutamine were related by their carbon metabolism through a common path involving glutamic acid. However, the main feature of glutamine metabolism was the removal of its α-amino and the amide nitrogen to yield α-keto acids especially α-ketoglutaric acid. The occurrence of α-ketoglutaramic acid could have accounted for succinamic acid and succinimide derived from 14C-L-glutamine.

1961 ◽  
Vol 39 (10) ◽  
pp. 1559-1566 ◽  
Author(s):  
H. Reisener ◽  
W. B. McConnell ◽  
G. A. Ledingham

When uredospores of Puccinia graminis var. tritici race 15B were shaken in a medium containing M/30 phosphate buffer, pH 6.2, and valerate-1-C14, 97% of the radioactivity was removed from the solution in a period of 3 hours. Fifty-five per cent of the carbon-14 was released as carbon dioxide, and 42% was incorporated into the spores. Carbon-14 was found in many cellular components but the water-soluble fraction accounted for 48% of the tracer in the spores. About two thirds of the water-soluble carbon-14 was in a fraction containing amino acids, amides, and peptides, with glutamic acid, glutamine, and γ-aminobutyric acid being highly radioactive. Carbon-5 of glutamic acid and carbon-1 of γ-aminobutyric acid were particularly radioactive. In addition carbon-1 of glutamic acid was appreciably radioactive. The results are consistent with the view that γ-aminobutyric acid was formed by decarboxylation of glutamic acid and that glutamic acid became labelled as a result of β-oxidation of the valerate-1-C14 to yield acetate-1-C14 which in turn was metabolized by the tricarboxylic acid cycle.


1965 ◽  
Vol 43 (1) ◽  
pp. 91-96 ◽  
Author(s):  
S. Suryanarayanan ◽  
W. B. McConnell

Uredospores of Puccinia graminis var. tritici were incubated in phosphate buffer (pH 6.2) containing pelargonic acid-1-C14. After 3 hours 97.5% of the tracer was assimilated. Fifty-five percent of this was released as C14O2 and 36.2% was incorporated into the spores. About one-half of the carbon-14 in the spores was soluble in ethanol and water, whereas nearly a third was ether extractable. The amino acid and carbohydrate fractions contained about equal amounts of carbon-14 and together accounted for two-thirds of the radioactivity in the ethanol–water extract. The organic acids were also radioactive. Glutamic acid, γ-aminobutyric acid, aspartic acid, and alanine were the most highly labelled amino acids. Fifty-three percent of the radioactivity in glutamic acid was found in carbon 1 and 46% in carbon 5. This distribution suggests β-oxidation of pelargonic acid to acetyl CoA and extensive utilization of the latter by means of the glyoxylate cycle.


1971 ◽  
Vol 49 (12) ◽  
pp. 2163-2173 ◽  
Author(s):  
D. J. Durzan ◽  
P. K. Ramaiah

The metabolism of L-proline was studied in 6-day-old jack pine seedlings, freshly excised from the nutritive female gametophyte. During the following 24 h, a sharp drop in free amino acids and protein was observed. Although levels of free proline were low, uniformly labeled 14C-L-proline and proline-3, 4-3H served as precursors for the dicarboxylic amino acids and their corresponding amides, glutamine and asparagine, which usually accumulate during germination. The origin of asparagine while unresolved did not involve β-cyanoalanine. Other products of proline metabolism included Δ1-pyrroline-5-carboxylic acid, glutamic acid, and γ-aminobutyric acid. With 14C-proline, radioactivity in alanine and serine resulted presumably from refixation of 14CO2 that was released by the decarboxylation of glutamic acid and other organic acids. The remaining products, e.g. pyrrolidone carboxylic acid, succinamic acid, and succinimide, were more closely related to the fate of glutamine than to proline.Radioactivity in proline and derived amino acids was recovered from soluble proteins separated on polyacrylamide gels. Five fractions revealed a similar diurnal turnover of specific activity. Three of these contained peroxidase isoenzyme activity. The recovery of tritium from peroxidase isoenzymes was related through the metabolism of proline to the intake and metabolism of water as well as to the appearance of enzyme activity in vascular tissues and emerging root and shoot apices.


1973 ◽  
Vol 51 (2) ◽  
pp. 351-358 ◽  
Author(s):  
D. J. Durzan

White spruce seedlings containing urease were exposed to 0.16 M urea for 4 h in continuous light. Seedlings accumulated total soluble nitrogen as amides and arginine, and increased their content of bound amino acid nitrogen. In darkness, total soluble nitrogen declined and the increase of total bound amino acids was not as great as in light. In both treatments, the fate of tritiated water was examined by recovery of nonexchangeable tritium from amino acids. As urea was consumed, more tritium was recovered from seedlings in light than in darkness. In both treatments tritium followed the nitrogen of urea and was bound covalently, initially in glutamic acid, and subsequently wherever an α-keto acid was a precursor for the synthesis of the corresponding amino acid, viz. alanine and aspartic acid. In light, tritium was recovered mainly from glutamic acid, followed by glutamine, and to a lesser extent by γ-aminobutyric acid. In darkness, while glutamic acid was prominent initially, more radioactivity was recovered from γ-aminobutyric and glutamic acids compared to glutamine and to the light treatment. Glutamic acid was the main bound amino acid containing covalent tritium.The occurrence of tritium at the α-carbon of glutamic acid was supported by transfer of this tritium after decarboxylation to γ-aminobutyric acid, and by conversion of bound glutamate-3H to radioactive pyrrolidone carboxylic acid during acid hydrolysis of protein.Although urea nitrogen contributed to arginine synthesis in light, no tritium was found in arginine nor its precursors in the ornithine cycle until later, when nearly all amino acids were radioactive. This is consistent with the absence of covalent binding of tritium in ureido precursors leading to arginine biosynthesis, and supports the idea that tritium did not readily follow the carbon of urea into covalent linkage.


2019 ◽  
Vol 25 (6) ◽  
pp. 785-791
Author(s):  
Woong-Kyu Yoon ◽  
Jae-Won Choi ◽  
Jong-Soon Lim ◽  
Coralia V. Garcia ◽  
Sam-Pin Lee

1950 ◽  
Vol 187 (1) ◽  
pp. 55-63 ◽  
Author(s):  
Eugene Roberts ◽  
Sam Frankel

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