scholarly journals Measurement of the rates of protein synthesis in rabbits. A method for the estimation of rates of change in the specific radioactivities of free amino acids during continuous infusions

1980 ◽  
Vol 192 (2) ◽  
pp. 623-629 ◽  
Author(s):  
G E Lobley ◽  
S P Robins ◽  
R M Palmer ◽  
I McDonald

1. A method is described by which, from analysis of terminal samples, the rate constants that define the changes in specific radioactivity of free amino acids during continuous infusions can be estimated. The method involves the infusion of 3H-labelled and 14C-labelled forms of an amino acid for different, but overlapping, periods. 2. The procedure was developed for infusions of proline and tyrosine into New Zealand White rabbits and the rate constants were determined for blood and muscle. 3. The rate constant for equilibrium of radioactive free proline was much lower in muscle than in blood, and indicated that a plateau condition may not be attained in muscle by the end of a 6 h infusion. 4. Comparison of the ratio of areas under the curves of estimated specific radioactivity plotted versus time with the 3H/14C ratio of bound amino acid in muscle protein suggested that radioactive proline and tyrosine may be incorporated preferentially from an extracellular rather than an intracellular source.

1964 ◽  
Vol 10 (5) ◽  
pp. 623-631 ◽  
Author(s):  
C. Ramirez ◽  
J. J. Miller

During 6-day exposures of cells of Saccharomyces cerevisiae to acetate sporulation medium, the content of free amino acids declined to approximately one-third of that of vegetative cells, but proline was exceptional in that it increased conspicuously in amount. The content of combined amino acids also diminished to about one-third, ammonia was evolved, and amino acids (not including proline) passed out of the cells into the medium. When dihydroxyacetone replaced acetate in the sporulation medium, the results were similar except that the decline in content of free and combined amino acids was much greater, more ammonia was evolved, and only very small amounts of amino acids could be detected in the medium. Transfer of sporulated cells to growth medium led to an increase in the pool of free amino acids, except for proline, which declined in amount.In two other species of Saccharomyces the free proline content also increased on exposure to sporulation medium, but in Schizosaccharomyces pombe and Torulopsis famata no such increase was observed.


1975 ◽  
Vol 147 (3) ◽  
pp. 473-477 ◽  
Author(s):  
M Nwagwu

A procedure for preparing polyribosome aminoacyl-tRNA free from contamination by supernatant aminoacyl-tRNA and free amino acids is described. Important features of the procedure are the use of acidic buffers to help protect the amino acid-tRNA linkage and the inclusion of sodium dodecyl sulphate, to inhibit ribonuclease activity. The specific radioactivity of polyribosome aminoacyl-tRNA is high within 30s and reaches a maximum in 2 1/2 min, well ahead of polyribosome peptides which, as described by Herrmann et al. (1971), attain maximum specific radioactivity in about 10 min.


1970 ◽  
Vol 48 (3) ◽  
pp. 228-235 ◽  
Author(s):  
Y. Yoshino ◽  
K. A. C. Elliott

The time course of entry of radioactive carbon from intravenously administered [U-14C]-glucose into free amino acids in the brains of rats has been studied using an automatic amino acid analyzer coupled through a flow cell with a scintillation counter. Radioactivity appeared rapidly in alanine, aspartic acid, glutamic acid, glutamine, and γ-aminobutyric acid as previously shown, and in an unknown ninhydrin-positive substance present in very small amount. Urea, serine, and glycine became slightly radioactive. Four hours after giving the radioactive glucose, the specific activity in all soluble substances was low. In pentobarbital anesthesia, specific radioactivity was increased in alanine and decreased in γ-aminobutyric acid, aspartic and glutamic acids, and glutamine. A high proportion of radioactivity remained in glucose. Under hypoxia, alanine increased in amount but decreased in specific activity, and the specific activities of the other strongly labelled amino acids decreased. The proportion of the total radioactivity found in glucose and lactate increased. During picrotoxin and pentylenetetrazol convulsions, changes occurred which were similar to those under hypoxia. After aminooxyacetic acid administration, the well-known great increase in γ-aminobutyric acid level was found to be accompanied by a decrease in glutamate and also in aspartic acid and alanine, indicating inhibition of the three transaminases concerned. The previously observed brief rapid postmortem increase in the amount of γ-aminobutyric acid was confirmed; alanine also increased briefly but no other amino acid did so. The increased γ-aminobutyric acid had the same specific radioactivity as the original but the extra alanine was less radioactive than the original. When the γ-aminobutyric acid level had been increased by administration of aminooxyacetic acid, the rapid postmortem increase did not occur.


1968 ◽  
Vol 46 (3) ◽  
pp. 217-223 ◽  
Author(s):  
R. M. Behki ◽  
S. M. Lesley

The concentration of each free amino acid present in hypocotyl tissue of sunflower seedlings was determined during the period following both sterile wounding and inoculation with either tumorigenic (strain 1025) or non-tumorigenic (strain IIBNV6) Agrobacterium tumefaciens. The concentration of the total free amino acids of hypocotyl tissue reached a maximum 68 hours after infection with strain 1025 and this level was 50% greater than that found in plants treated with strain IIBNV6. The total content of free basic amino acids increased to a peak value at 20 hours after infection by strain 1025 and this represented 1.4 to 3 times the level found in strain IIBNV6 inoculated seedlings at 20 hours.The amount of free proline decreased with time and it was essentially absent 164 hours after infection with strain 1025. At 2 months the concentration of free proline in the tumor tissue was greatly increased with 75% of the total free amino acids accounted for as proline.The free amino acid content of strain 1025 growing in either the presence or absence of an extract from wounded seedlings was measured and the results indicated no significant relationship between the plant and bacterial systems with regard to their free amino acid pools.


2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 673-673
Author(s):  
Michelle E G Weijzen ◽  
Rob JJ van Gassel ◽  
Imre W K Kouw ◽  
Stefan H M Gorissen ◽  
Marcel CG van de Poll ◽  
...  

Abstract Objectives The rate of protein digestion and amino acid absorption determines the postprandial rise in circulating amino acids and, as such, modulates postprandial muscle protein synthesis rates. This study compares protein digestion and amino acid absorption kinetics and the subsequent muscle protein synthetic response following ingestion of intact protein versus an equivalent amount of free, crystalline amino acids. Methods Twenty-four healthy, young subjects (age: 22 ± 3 y, BMI: 23 ± 2 kg·m−2, sex: 12 M/12F) ingested 30 g intrinsically L-[1–13C]-phenylalanine and L-[1–13C]-leucine labeled milk protein (PROT; n = 12) or an equivalent amount of free amino acids (AA; n = 12). In addition, subjects received primed continuous L-[ring-2H5]-phenylalanine, L-[ring-3,5–2H2]-tyrosine, and L-[1–13C]-leucine infusions. Blood samples and muscle biopsies were obtained frequently to assess protein digestion and amino acid absorption kinetics and subsequent muscle protein synthesis rates over a 6 h postprandial period. An unpaired t-test was used to compare overall exogenous phenylalanine release in plasma. For other parameters repeated measures ANOVA were applied to determine differences between groups over time (time as within, and group as between-subjects factor). Data are expressed as mean ± SD. Results Postprandial plasma amino acid concentrations and exogenous phenylalanine appearance rates increased after ingestion of PROT and AA (both, P < 0.001), with a greater increase following ingestion of AA when compared to PROT (time*group interaction P < 0.001). Exogenous phenylalanine release in plasma assessed over the 6 h postprandial period, was greater in AA (76 ± 9%) compared with PROT (59 ± 10%; P < 0.001). Ingestion of AA and PROT strongly increased muscle protein synthesis rates based upon L-[ring-2H5]-phenylalanine (time effect P < 0.001), with no differences between groups (from 0.037 ± 0.015 to 0.053 ± 0.014%·h−1 and from 0.039 ± 0.016 to 0.051 ± 0.010%·h−1, respectively; time*group interaction P = 0.629). Conclusions Ingestion of free amino acids as opposed to intact milk protein is followed by more rapid amino acid absorption and greater postprandial plasma amino acid availability, but this does not further augment postprandial muscle protein synthesis rates. Funding Sources This research did not receive external funding.


1973 ◽  
Vol 28 (7-8) ◽  
pp. 449-451 ◽  
Author(s):  
G. Peter ◽  
H. Angst ◽  
U. Koch

Free and protein-bound amino acids in serum and scales were investigated. In serum the bound amino acids of psoriatics are significantly higher with exception of Pro, Met, Tyr and Phe in contrast to normal subjects. For free amino acids the differences between normal subjects and psoriatics found in serum and scales are not significant. Results are discussed in relation to the single amino acids and the biochemical correlations are outlined which takes the pathological process as a basis.


1978 ◽  
Vol 77 (1) ◽  
pp. 15-31 ◽  
Author(s):  
R. K. O'DOR ◽  
M. J. WELLS

Octopus vulgaris can be forced into precocious maturity by removal of the subpedunculate lobe from the brain, an operation that releases the optic glands from inhibition, and allows them to secrete a gonadotropin. 14C-leucine was injected into the bloodstream of immature animals and its subsequent incorporation into muscle protein followed by taking successive samples from the arms. The optic glands were then activated, and a further injection of 3H-leucine given and followed by means of further arm samples. Optic gland secretion suppresses protein synthesis in the muscles. This is associated with an increase in the total amino acid pool in the muscles and with a considerable increase in the concentration of free amino acids circulating in the blood. If an ovary is present these events are associated with a rapid growth of the ovary and its ducts, and a loss of weight elsewhere. In ovariectomized animals the ducts grow, but there is no yolk to absorb the large pool of free amino acids, and the animals gain weight by osmotic uptake of water into the muscles. The developing ovary may produce a hormone that increases the release of amino acids from muscle, since the concentration circulating in the blood of intact animals remains at least as high as in ovariectomized octopuses, despite the demands of the developing ovary. These matters are discussed in relation to other evidence for a gonadial hormone and in relation to the ‘self-destruct’ effect of the optic gland secretion in determining the post-reproductive death of octopuses.


1980 ◽  
Vol 239 (6) ◽  
pp. G493-G496 ◽  
Author(s):  
E. J. Feldman ◽  
M. I. Grossman

Using intragastric titration in dogs with gastric fistulas, dose-response studies were carried out with liver extract and with a mixture of amino acids that matched the free amino acids found in liver extract. All solutions were adjusted to pH 7.0 and osmolality to 290 mosmol x kg-1. Doses are expressed as the sum of the concentrations of all free amino acids. At each dose studied (free amino acid concentration: 2.8, 5.6, 11, 23, and 45 mM), acid secretion in response to the free amino acid mixture was not significantly different from that of liver extract. The peak response to both liver extract and the free amino acid mixture occurred with the 23-mM dose and represented about 60% of the maximal response to histamine. The serum concentrations of gastrin after liver extract and the amino acid mixture were not significantly different. It is concluded that in dogs with gastric fistula, gastric acid secretion and release of gastrin were not significantly different in response to liver extract and to a mixture of amino acids that simulated the free amino acid content of liver extract.


1968 ◽  
Vol 96 (3) ◽  
pp. 303-318 ◽  
Author(s):  
P. M.-B. Leung ◽  
Q. R. Rogers ◽  
A. E. Harper

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