Proteomic Profiling of De Novo Protein Synthesis in Starvation-Induced Autophagy Using Bioorthogonal Noncanonical Amino Acid Tagging

2017 ◽  
pp. 41-59 ◽  
Author(s):  
J. Zhang ◽  
J. Wang ◽  
Y.-M. Lee ◽  
T.-K. Lim ◽  
Q. Lin ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
De Novo ◽  
Proteomic Profiling ◽  
Noncanonical Amino Acid ◽  
De Novo Protein Synthesis

Transient increase of de novo amino acid synthesis and its physiological significance in water-stressed white clover

2005 ◽  
Vol 32 (9) ◽  
pp. 831 ◽  
Author(s):  
Bok-Rye Lee ◽  
Woo-Jin Jung ◽  
Kil-Yong Kim ◽  
Jean-Christophe Avice ◽  
Alain Ourry ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Water Deficit ◽  
White Clover ◽  
De Novo ◽  
Acid Synthesis ◽  
Ammonia Concentration ◽  
De Novo Synthesis ◽  
Amino Acid Synthesis ◽  
De Novo Protein Synthesis

In white clover (Trifolium repens L. cv. Regal) the kinetics of de novo synthesis of amino acid and protein were compared by tracing 15N under well-watered (control) or water-deficit conditions. The physiological relationship between ammonia concentration, in response to the change in leaf water parameters, and de novo synthesis of amino acid and protein was also assessed. Leaf and root dry mass were not significantly affected for the first 3 d, whereas metabolic parameters such as total N and ammonia were significantly affected within the first day of water-deficit treatment. Inhibitory effect of water deficit on N acquisition from the soil was significant throughout the experimental period. Water deficit induced a significant increase in ammonia concentration in leaves during the first 3 d, and in roots for only the first day. In both leaves and roots, an increase in de novo amino acid synthesis, which peaked in leaves within the first 3 d of water-deficit treatment (Ψw ≥ –1.18 MPa), was observed. The rate of decrease in de novo protein synthesis gradually accelerated as the duration of the water-deficit treatment increased. There was a significant positive relationship between ammonia production and the increase in de novo amino acid synthesis during the first 3-d period, but not during the later period (day 3–day 7). This experiment clearly indicates that the increase in de novo amino acid synthesis caused by water deficit is a transient adaptive response occurring during the first few days and that it is associated with the increased ammonia concentrations, which in turn arise in response to a decrease in de novo protein synthesis.

scholarly journals Development, amino acid utilization and cell allocation in bovine embryos after in vitro production in contrasting culture systems

Reproduction ◽  
2002 ◽  
pp. 155-165 ◽  
Author(s):  
M Kuran ◽  
JJ Robinson ◽  
DS Brown ◽  
TG McEvoy
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
De Novo ◽  
Acid Uptake ◽  
Free Medium ◽  
Cell Counts ◽  
De Novo Protein Synthesis ◽  
Amino Acid Flux ◽  
Protein Free Medium

The effects of protein-supplemented and protein-free media on amino acid uptake, protein synthesis and cell differentiation in bovine blastocysts were investigated. Four formulations of synthetic oviduct fluid were used. Each formulation was identified by the principal supplement: bovine serum albumin (0.4%, w/v); polyvinyl alcohol (0.3%, w/v); or either of two steer sera (10%, v/v). After zygote culture, blastocyst yields (day 7.5) were lowest in protein-free medium and highest in albumin-supplemented medium. Subsequent 12 h incubation in the presence of both essential and non-essential amino acids was used for the measurement of amino acid flux. All blastocysts released alanine but consumed aspartate (P < 0.001) and the extent was influenced by prior culture conditions. Aspartate uptake was lower in blastocysts produced in protein-free conditions (P < 0.05) than in blastocysts produced in albumin-supplemented conditions. Consumption indices for 16 other amino acids were not influenced by blastocyst source. Cell counts and hatching incidences were highest for albumin-supplemented blastocysts, but were similar among blastocysts from the protein-free and serum-dependent treatments. Crucially, the use of protein-free medium for zygote culture did not compromise resultant blastocysts in terms of either de novo protein synthesis ([3H]phenylalanine incorporation) or trophectoderm function (phenotype based on interferon-tau detection). Thus, although blastocyst yields were compromised after zygote culture in a protein-free (vis-a-vis albumin-supplemented) medium, amino acid flux was qualitatively conserved, and only quantitatively modified in the case of alanine and aspartate. Moreover, vital properties of blastocysts that were produced, including de novo protein synthesis and trophectodermal cell function, apparently were not adversely affected by protein deprivation.

Amino Acid Transport into Human Platelets and Subsequent Incorporation into Protein

1970 ◽  
Vol 23 (01) ◽  
pp. 140-147 ◽  
Author(s):  
I. A Cooper ◽  
B. G Firkin
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Amino Acid Transport ◽  
De Novo ◽  
Human Platelets ◽  
Acid Transport ◽  
Protein Components ◽  
De Novo Protein Synthesis ◽  
Subsequent Incorporation

SummaryHuman platelets are known to contain various protein components. Among them are fibrinogen and other soluble proteins. The origin of such proteins has not been clear.Studies were designed to demonstrate the ability of the platelet to incorporate amino acids and subsequently utilise these for de novo protein synthesis.Seven different 14C labelled amino acids were used, cysteine, lysine, serine, glutamic acid, proline, valine und leucine.Active incorporation into platelets of all these amino acids was demonstrated with some evidence for incorporation of cysteine, lysine and serine into the fibrinogen and soluble protein.

scholarly journals Viral Gene Expression Patterns in Human Herpesvirus 6B-Infected T Cells

2002 ◽  
Vol 76 (15) ◽  
pp. 7578-7586 ◽  
Author(s):  
Bodil Øster ◽  
Per Höllsberg
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
Protein Synthesis ◽  
De Novo ◽  
Expression Patterns ◽  
Human Herpesvirus ◽  
Viral Gene Expression ◽  
Polymerase Activity ◽  
Viral Gene ◽  
De Novo Protein Synthesis

ABSTRACT Herpesvirus gene expression is divided into immediate-early (IE) or α genes, early (E) or β genes, and late (L) or γ genes on the basis of temporal expression and dependency on other gene products. By using real-time PCR, we have investigated the expression of 35 human herpesvirus 6B (HHV-6B) genes in T cells infected by strain PL-1. Kinetic analysis and dependency on de novo protein synthesis and viral DNA polymerase activity suggest that the HHV-6B genes segregate into six separate kinetic groups. The genes expressed early (groups I and II) and late (groups V and VI) corresponded well with IE and L genes, whereas the intermediate groups III and IV contained E and L genes. Although HHV-6B has characteristics similar to those of other roseoloviruses in its overall gene regulation, we detected three B-variant-specific IE genes. Moreover, genes that were independent of de novo protein synthesis clustered in an area of the viral genome that has the lowest identity to the HHV-6A variant. The organization of IE genes in an area of the genome that differs from that of HHV-6A underscores the distinct differences between HHV-6B and HHV-6A and may provide a basis for further molecular and immunological analyses to elucidate their different biological behaviors.

De novo protein synthesis in relation to ammonia and proline accumulation in water stressed white clover

2004 ◽  
Vol 31 (8) ◽  
pp. 847 ◽  
Author(s):  
Tae-Hwan Kim ◽  
Bok-Rye Lee ◽  
Woo-Jin Jung ◽  
Kil-Yong Kim ◽  
Jean-Christophe Avice ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Water Deficit ◽  
White Clover ◽  
De Novo ◽  
Proline Accumulation ◽  
Leaf Water ◽  
Total N ◽  
Water Parameters ◽  
Leaves And Roots ◽  
De Novo Protein Synthesis

The kinetics of protein incorporation from newly-absorbed nitrogen (N, de novo protein synthesis) was estimated by 15N tracing in 18-week-old white clover plants (Trifolium repens L. cv. Regal) during 7 d of water-deficit treatment. The physiological relationship between kinetics and accumulation of proline and ammonia in response to the change in leaf-water parameters was also assessed. All leaf-water parameters measured decreased gradually under water deficit. Leaf and root dry mass was not significantly affected during the first 3 d when decreases in leaf-water parameters were substantial. However, metabolic parameters such as total N, proline and ammonia were significantly affected within 1 d of commencement of water-deficit treatment. Water-deficit treatment significantly increased the proline and NH3–NH4+ concentrations in both leaves and roots. There was a marked reduction in the amount of N incorporated into the protein fraction from the newly absorbed N (NANP) in water-deficit stressed plants, particularly in leaf tissue. This reduction in NANP was strongly associated with an increased concentration of NH3–NH4+ in roots (P≤0.05) and proline (P≤0.01) in leaves and roots. These results suggest that proline accumulation may be a sensitive biochemical indicator of plant water status and of the dynamics of de novo protein synthesis in response to stress severity.

Growth hormone decreases muscle glutamine production and stimulates protein synthesis in hypercatabolic patients

2000 ◽  
Vol 279 (2) ◽  
pp. E323-E332 ◽  
Author(s):  
Gianni Biolo ◽  
Fulvio Iscra ◽  
Alessandra Bosutti ◽  
Gabriele Toigo ◽  
Beniamino Ciocchi ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Protein Synthesis ◽  
Amino Acid ◽  
De Novo ◽  
Muscle Protein ◽  
Recombinant Human Growth Hormone ◽  
Glutamine Metabolism ◽  
Trauma Patients ◽  
Mrna Levels ◽  
Glutamine Production

We determined the effects of 24-h recombinant human growth hormone (rhGH) infusion into a femoral artery on leg muscle protein kinetics, amino acid transport, and glutamine metabolism in eight adult hypercatabolic trauma patients. Metabolic pathways were assessed by leg arteriovenous catheterization and muscle biopsies with the use of stable amino acid isotopes. Muscle mRNA levels of selected enzymes were determined by competitive PCR. rhGH infusion significantly accelerated the inward transport rates of phenylalanine and leucine and protein synthesis, whereas the muscle protein degradation rate and cathepsin B and UbB polyubiquitin mRNA levels were not significantly modified by rhGH. rhGH infusion decreased the rate of glutamine de novo synthesis and glutamine precursor availability, total branched-chain amino acid catabolism, and nonprotein glutamate utilization. Thus net glutamine release from muscle into circulation significantly decreased after rhGH administration (∼50%), whereas glutamine synthetase mRNA levels increased after rhGH infusion, possibly to compensate for reduced glutamine precursor availability. We conclude that, after trauma, the anticatabolic action of rhGH is associated with a potentially harmful decrease in muscle glutamine production.

scholarly journals Action of growth hormone in vitro on the net uptake and incorporation into protein of amino acids in muscle from intact rabbits given protein-deficient diets

1976 ◽  
Vol 35 (1) ◽  
pp. 1-10 ◽  
Author(s):  
M. R. Turner ◽  
P. J. Reeds ◽  
K. A. Munday
Keyword(s):  
Amino Acids ◽  
Growth Hormone ◽  
Protein Synthesis ◽  
Amino Acid ◽  
De Novo ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
Amino Acid Incorporation ◽  
Acid Incorporation

1. Net amino acid uptake, and incorporation into protein have been measured in vitro in the presence and absence of porcine growth hormone (GH) in muscle from intact rabbits fed for 5 d on low-protein (LP), protein-free (PF) or control diets.2. In muscle from control and LP animals GH had no effect on the net amino acid uptake but stimulated amino acid incorporation into protein, although this response was less in LP animals than in control animals.3. In muscle from PF animals, GH stimulated both amino acid incorporation into protein and the net amino acid uptake, a type of response which also occurs in hypophysectomized animals. The magnitude of the effect of GH on the incorporation of amino acids into protein was reduced in muscle from PF animals.4. The effect of GH on the net amino acid uptake in PF animals was completely blocked by cycloheximide; the uptake effect of GH in these animals was dependent therefore on de novo protein synthesis.5. It is proposed that in the adult the role of growth hormone in protein metabolism is to sustain cellular protein synthesis when there is a decrease in the level of substrate amino acids, similar to that which occurs during a short-term fast or when the dietary protein intake is inadequate.

Sign in / Sign up

Export Citation Format

Share Document