A note on the incorporation of amino acids by hamster embryo cells in monolayer culture

Taylor, Joyce; Stanners, Clifford P.

doi:10.1016/0005-2787(67)90566-7

Cited by 7 publications

(5 citation statements)

References 4 publications

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“…Clark & Schmidt, 1967;Taylor & Stanners, 1967). In both preparations, a greater proportion of the amino acid taken up was present in the acid-soluble fraction when the higher external concentration of the amino acids was used.…”

Section: Discussionmentioning

confidence: 95%

“…The amount of an amino acid taken up by mammalian cells increases with an increase in its extracellular concentration within a wide range (inter alia, Clark & Schmidt, 1967;Taylor & Stanners, 1967); therefore, for any comparison of the ability of different cell systems to take up amino acids to be valid, it would be advisable to study the uptake at at least two (preferably more, to allow kinetic analysis) concentrations, one low (as in Table 1) and the other high. For this reason, we also studied the total uptake of four amino acids (leucine, valine, lysine and arginine) in normal liver, in a solid hepatoma and in the Zajdela ascitic hepatoma cells, in a serum-free tissue culture medium in which the above amino acids were present at concentrations varying from 0.8 to 1.2 raM, that is, about a 1 000-fold higher than in Table 1.…”

Section: Uptake Of Amino Acids At High Extracellular Concentrationsmentioning

confidence: 98%

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A comparison of the ability of normal liver, a premalignant liver, a solid hepatoma and the zajdela ascitic hepatoma, to take up amino acidsin vitro

et al. 1976

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The net total uptake of several amino acids at low (0.8-3.1 mumoles/liter) as well as high (800-1200 mumoles/liter) extracellular concentrations, by normal rat liver, a premalignant liver, a solid hepatoma, and the Zajdela ascitic hepatoma cells, has been compared under conditions in which protein synthesis continues. At low amino acid concentrations, the initial (3 min) total uptake of the various amino acids in the Zajdela cells, was 3-10 (average 7) times more, and the intracellular concentration of the labeled amino acids taken up 14-45 (average 31) times more, than in normal liver. At the high amino acid concentrations, the total uptake in the Zajdela cells, at 60-120 min was 2-5 (average 3.5) times more, and the intracellular concentration of the amino acids taken up 8-18 (average 13) times more, than in normal liver; the corresponding values for the premalignant liver and the solid hepatoma were in between those for normal liver and the Zajdela cells. Further, the rate of the total uptake of amino acids, their intracellular concentration, the proportion of the amino acid taken up utilized for protein synthesis, the rate of incorporation of the amino acid taken up into protein, and the cellular growth rate, seemed to be correlated in the four cell/tissue preparations studied. In most cases, the rate of the net uptake fell drastically with time, the uptake virtually stopping after 90-180 min, probably due to lack of serum in the incubation medium.

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Section: Discussionmentioning

confidence: 95%

Section: Uptake Of Amino Acids At High Extracellular Concentrationsmentioning

confidence: 98%

A comparison of the ability of normal liver, a premalignant liver, a solid hepatoma and the zajdela ascitic hepatoma, to take up amino acidsin vitro

et al. 1976

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“…After exactly 30 min, 0.5 ml of a solution of 14C-valine was added to the growth medium to give a final concentration of 0.87 ,uc/ml in experiment 1, and 2 ,uc/ml in the other three experiments. It should be stressed at this point that the absolute valine concentration in the medium (47 jig/ml) was not reduced, as such treatment has been found to reduce the rate of protein synthesis in these cells rapidly and permanently (Taylor and Stanners, 1967).…”

Section: Solutions and Chemicalsmentioning

confidence: 95%

“…5 that (P + S)' became constant, and therefore, the valine pool reached its maximum specific activity within 5 min after addition of labelled valine. Since the cells would require about 1 to 2 hr to incorporate all the valine present in the intracellular pool into protein (Taylor, 1967),3 the valine pool must become labelled by a rapid exchange of valine between cells and medium (Britten and McClure, 1962), rather than by uptake of valine to supply the need for protein synthesis. Also the size of the total cellular valine pool is small compared to the total amount of valine in the medium (Taylor, 1967).…”

Section: The Simplest Casementioning

confidence: 99%

“…Since the cells would require about 1 to 2 hr to incorporate all the valine present in the intracellular pool into protein (Taylor, 1967),3 the valine pool must become labelled by a rapid exchange of valine between cells and medium (Britten and McClure, 1962), rather than by uptake of valine to supply the need for protein synthesis. Also the size of the total cellular valine pool is small compared to the total amount of valine in the medium (Taylor, 1967). If the final specific activity of the cells plus medium is b cpm/mole, the total pool size is constant at d mole, and the total exchange rate is a mole/sec, then the rate of change of B(t) is given by:…”

Section: The Simplest Casementioning

confidence: 99%

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Polyribosomes of Hamster Cells

Stanners¹

1968

Biophysical Journal

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The amount of radioactivity incorporated into completed polypeptides and into nascent polypeptides bound to polyribosomes was measured for rapidly dividing, cultured hamster embryo cells incubated in the presence of labelled amino acids. Using a mathematical analysis which took into account variations in the specific activity of the intracellular amino acid pool, these measurements yielded a value for the length of time required for the synthesis of a primary polypeptide of average length, the "transit time." Subject to a number of reasonable assumptions in the mathematical analysis which were, however, only approximately verified, the transit time was found to be 23 +/- 2 sec in four independent experiments. Regardless of the absolute accuracy of this measurement, the method is considered a useful measure of translation independent of transcription.

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Control of macromolecular synthesis in proliferating and resting syrian hamster cells in monolayer culture. III. Electrophoretic patterns of newly synthesized proteins in synchronized proliferating cells and resting cells

Becker

Stanners

1972

Journal Cellular Physiology

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Electrophoretic patterns of newly synthesized proteins have been compared for hamster embryo fibroblasts in asynchronous cultures, mitotically synchronized cultures, and stationaq phase cultures. Only proteins with molecular weight between 30,000 and 150,000, comprising 60-70 7i of the total cell proteins and excluding histones and collagen were included in the comparison. Although no significant differences could be detected between such patterns for cells at different stages of the cell cycle, significant differences were detectcd between patterns for cells i n stationary phase and for proliferating asynchronous or synchronous cells in any stage of the cell cycle. These differences amounted to at least 5%) of the newly synthesized cellular proteins. Much larger differences were detected between patterns €rom a nuclear fraction of proliferating and resting cells. These results indicate that normal cells in stationary phase are arrested in a state distinct from any phase of the normal cell cycle and may provide a biochemical marker for resting cells.

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A note on the incorporation of amino acids by hamster embryo cells in monolayer culture

Cited by 7 publications

References 4 publications

A comparison of the ability of normal liver, a premalignant liver, a solid hepatoma and the zajdela ascitic hepatoma, to take up amino acidsin vitro

A comparison of the ability of normal liver, a premalignant liver, a solid hepatoma and the zajdela ascitic hepatoma, to take up amino acidsin vitro

Polyribosomes of Hamster Cells

Control of macromolecular synthesis in proliferating and resting syrian hamster cells in monolayer culture. III. Electrophoretic patterns of newly synthesized proteins in synchronized proliferating cells and resting cells

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