scholarly journals Protein turnover in the developing Triticum aestivum grain

2021 ◽  
Author(s):  
Hui Cao ◽  
Owen Duncan ◽  
A. Harvey Millar
Keyword(s):  
Protein Synthesis ◽  
Protein Turnover ◽  
Storage Proteins ◽  
Spatiotemporal Pattern ◽  
Turnover Rates ◽  
Grain Development ◽  
Wheat Grain ◽  
Stable Isotope Labelling ◽  
Atp Production

Protein abundance in cereal grains is determined by the relative rates of protein synthesis and protein degradation during grain development. Through combining in vivo stable isotope labelling and in-depth quantitative proteomics, we have measured the turnover of 1400 different types of proteins during wheat grain development. We demonstrate that there is a spatiotemporal pattern to protein turnover rates which explain part of the variation in protein abundances that is not attributable to differences in wheat gene expression. We show that approximately 20% of total grain ATP production is used for grain proteome biogenesis and maintenance, and nearly half of this budget is invested exclusively in storage protein synthesis. We calculate that 25% of newly synthesized storage proteins are turned over during grain development rather than stored. This approach to measure protein turnover rates at proteome scale reveals how different functional categories of grain proteins accumulate, calculates the costs of protein turnover during wheat grain development and identifies the most and the least stable proteins in the developing wheat grain.

scholarly journals Protein turnover and growth in the whole body, liver and kidney of the rat from the foetus to senility

1984 ◽  
Vol 217 (2) ◽  
pp. 507-516 ◽  
Author(s):  
D F Goldspink ◽  
F J Kelly
Keyword(s):  
Protein Synthesis ◽  
Protein Turnover ◽  
Post Partum ◽  
Protein Composition ◽  
Whole Body ◽  
Turnover Rates ◽  
Tissue Mass ◽  
Age Related ◽  
Foetal Life

Changes in the growth and protein turnover (measured in vivo) of the rat liver, kidney and whole body were studied between 16 days of life in utero and 105 weeks post partum. Tissue and whole-body growth were related to changes in both cellular hyperplasia (i.e. changes in DNA) and hypertrophy (protein/DNA values) and to the protein composition within the enlarging tissue mass. The suitability of using a single large dose of phenylalanine for measuring the rates of protein synthesis during both pre- and post-natal life was established. The declining growth rates in the whole animal and the two visceral tissues were then explained by developmental changes in the fractional rates of protein synthesis and breakdown, turnover rates being age-for-age higher in the liver than in the kidney, which in turn were higher than those measured in the whole animal. The declining fractional rates of synthesis in both tissues and the whole body with increasing age were related to changes in the tissues' ribosomal capacity and activity. The fall in the hepatic rate between 18 and 20 days of foetal life (from 134 to 98% per day) corresponded to a decrease in both the ribosomal capacity and the rate of synthesis per ribosome. No significant changes in any of these parameters were, however, found in the liver between weaning (3 weeks) and senility (105 weeks). In contrast, the fractional synthetic (and degradative) rates progressively declined in the kidney (from 95 to 24% per day) and whole body (from 70 to 11% per day) throughout both pre- and post-natal life, mainly as a consequence of a progressive decline in the ribosomal capacity, but with some fall in the ribosomal activity also during foetal life. The age-related contributions of these visceral tissues to the total amount of protein synthesized per day by the whole animal were determined. The renal contribution remained fairly constant at 1.6-2.9%, whereas the hepatic contribution declined from 56 to 11%, with increasing age. Approximate-steady-state conditions were reached at, and between, 44 and 105 weeks post partum, the half-life values of mixed whole-body, kidney and liver proteins being 6.4, 3.0 and 1.5 days, respectively, at 105 weeks.

Respiratory energy requirements and rate of protein turnover in vivo determined by the use of an inhibitor of protein synthesis and a probe to assess its effect

1994 ◽  
Vol 92 (4) ◽  
pp. 585-594 ◽  
Author(s):  
T. J. Bouma ◽  
R. De Visser ◽  
J. H. J. A. Janssen ◽  
M. J. De Kock ◽  
P H. Van Leeuwen ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Protein Turnover ◽  
Energy Requirements ◽  
Inhibitor Of Protein Synthesis

scholarly journals The effect of protein depletion and repletion on muscle-protein turnover in the chick

1981 ◽  
Vol 194 (3) ◽  
pp. 811-819 ◽  
Author(s):  
M L MacDonald ◽  
R W Swick
Keyword(s):  
Protein Synthesis ◽  
Protein Turnover ◽  
Muscle Protein ◽  
Control Diet ◽  
Breast Muscle ◽  
Synthesis Rate ◽  
Protein Depletion ◽  
Protein Mass ◽  
Fractional Rate

Rates of growth and protein turnover in the breast muscle of young chicks were measured in order to assess the roles of protein synthesis and degradation in the regulation of muscle mass. Rates of protein synthesis were measured in vivo by injecting a massive dose of L-[1-14C]valine, and rates of protein degradation were estimated as the difference between the synthesis rate and the growth rate of muscle protein. In chicks fed on a control diet for up to 7 weeks of age, the fractional rate of synthesis decreased from 1 to 2 weeks of age and then changed insignificantly from 2 to 7 weeks of age, whereas DNA activity was constant for 1 to 7 weeks. When 4-week-old chicks were fed on a protein-free diet for 17 days, the total amount of breast-muscle protein synthesized and degraded per day and the amount of protein synthesized per unit of DNA decreased. Protein was lost owing to a greater decrease in the rate of protein synthesis, as a result of the loss of RNA and a lowered RNA activity. When depleted chicks were re-fed the control diet, rapid growth was achieved by a doubling of the fractional synthesis rate by 2 days. Initially, this was a result of increased RNA activity; by 5 days, the RNA/DNA ratio also increased. There was no evidence of a decrease in the fractional degradation rate during re-feeding. These results indicate that dietary-protein depletion and repletion cause changes in breast-muscle protein mass primarily through changes in the rate of protein synthesis.

Fiber-type composition of nine rat muscles. II. Relationship to protein turnover

1989 ◽  
Vol 257 (6) ◽  
pp. E828-E832 ◽  
Author(s):  
P. J. Garlick ◽  
C. A. Maltin ◽  
A. G. Baillie ◽  
M. I. Delday ◽  
D. A. Grubb
Keyword(s):  
Protein Synthesis ◽  
Regression Analysis ◽  
Protein Turnover ◽  
Fiber Type ◽  
Female Rats ◽  
Protein Breakdown ◽  
Fiber Type Composition ◽  
Type Composition ◽  
Age Regression

Rates of protein synthesis in vivo and fiber-type composition were measured in nine limb muscles of female rats at ages ranging from weaning to 1 yr. In all muscles, there was a decline in protein synthesis with increasing age, mostly as a result of a fall in the RNA content. Rates of protein breakdown and growth were determined in six muscles and these also declined with age. Regression analysis of the data for all ages showed that protein synthesis was correlated with the content of slow oxidative fibers but not with the relative proportions of fast glycolytic to fast oxidative glycolytic fibers.

scholarly journals Protein turnover rates of two human subjects during an unassisted crossing of Antarctica

1996 ◽  
Vol 76 (2) ◽  
pp. 165-174 ◽  
Author(s):  
M. A. Stroud ◽  
A. A. Jackson ◽  
J. C. Waterlow
Keyword(s):  
Protein Synthesis ◽  
Protein Turnover ◽  
Human Subjects ◽  
Austral Summer ◽  
High Energy ◽  
Turnover Rates ◽  
Energy Expenditures ◽  
Combined Stresses ◽  
Freeze Dried ◽  
Product Method

During the Austral summer of 1992–3, two men, MS and RF, walked 2300 km across Antarctica in 96 d, unassisted by other men, animals or machines. During the journey they ate freeze-dried rations, towed on sledges, that contained an average of 21·3 MJ/d of which 56·7% was fat, 35·5% carbohydrate and 7·8% protein (98·8 g). Despite this high energy intake both men lost more than 20 kg in body weight due to their extremely high energy expenditures. Studies of protein turnover using [15N]glycine by the single-dose end-product method were made before, during and after the journey, and these demonstrated considerable differences in the metabolic responses of the two men to the combined stresses of exercise, cold and undernutrition. However, both men maintained high and relatively stable levels of protein synthesis during the expedition despite the great exertion and the onset of considerable debilitation. This stability indicates the vital physiological function of protein synthesis.

Alterations of protein turnover underlying disuse atrophy in human skeletal muscle

2009 ◽  
Vol 107 (3) ◽  
pp. 645-654 ◽  
Author(s):  
S. M. Phillips ◽  
E. I. Glover ◽  
M. J. Rennie
Keyword(s):  
Protein Synthesis ◽  
Muscle Mass ◽  
Protein Turnover ◽  
Cultured Cells ◽  
Muscle Protein ◽  
Human Muscle ◽  
Muscle Loss ◽  
Disuse Atrophy ◽  
In Vivo Measurements

Unloading-induced atrophy is a relatively uncomplicated form of muscle loss, dependent almost solely on the loss of mechanical input, whereas in disease states associated with inflammation (cancer cachexia, AIDS, burns, sepsis, and uremia), there is a procatabolic hormonal and cytokine environment. It is therefore predictable that muscle loss mainly due to disuse alone would be governed by mechanisms somewhat differently from those in inflammatory states. We suggest that in vivo measurements made in human subjects using arterial-venous balance, tracer dilution, and tracer incorporation are dynamic and thus robust by comparison with static measurements of mRNA abundance and protein expression and/or phosphorylation in human muscle. In addition, measurements made with cultured cells or in animal models, all of which have often been used to infer alterations of protein turnover, appear to be different from results obtained in immobilized human muscle in vivo. In vivo measurements of human muscle protein turnover in disuse show that the primary variable that changes facilitating the loss of muscle mass is protein synthesis, which is reduced in both the postabsorptive and postprandial states; muscle proteolysis itself appears not to be elevated. The depressed postprandial protein synthetic response (a phenomenon we term “anabolic resistance”) may even be accompanied by a diminished suppression of proteolysis. We therefore propose that most of the loss of muscle mass during disuse atrophy can be accounted for by a depression in the rate of protein synthesis. Thus the normal diurnal fasted-to-fed cycle of protein balance is disrupted and, by default, proteolysis becomes dominant but is not enhanced.

scholarly journals PROTEIN METABOLISM IN TUMOR CELLS AT VARIOUS STAGES OF GROWTH IN VIVO

1974 ◽  
Vol 62 (3) ◽  
pp. 585-593 ◽  
Author(s):  
Massimo Olivotto ◽  
Francesco Paoletti
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Tumor Cells ◽  
Protein Metabolism ◽  
Protein Turnover ◽  
Specific Activity ◽  
Cell Mass ◽  
Ascites Hepatoma ◽  
Two Phases

Protein metabolism of Yoshida ascites hepatoma cells was studied in the early phase of logarithmic proliferation and in the following stage in which cell mass remains constant (resting phase). The rate of protein synthesis was measured by a short-time incorporation of [8H]lysine, while degradation was concurrently assessed by following the decrease of specific activity of [14C]lysine-labeled proteins. Most of the labeled amino acid injected intraperitoneally into the animal was immediately available for the tumor cells, with only a minor loss towards the extra-ascitic compartment. It was thus possible to calculate the dilution of the isotope in the ascitic pool of the lysine, which increased concurrently with the ascitic plasma volume. Amino acid transport capacity did not change in the log vs. the resting cells. This fact permitted the correction of the specific activity of the proteins synthesized by tumors in the two phases, taking into account the dilution effect. Protein synthesis was found to proceed at a constant rate throughout each of the two phases, although it was 30% lower during the resting as compared to the log phase. When cell mass attained the steady-state, protein degradation occurred at such a level as to balance the synthesis. Throughout the resting phase the amount of lysine taken up by the cells and renewed from the blood remained unchanged. Protein turnover, as studied in subcellular fractions, exhibited a similar rate in nuclei and microsomes, where it proceeded at a higher level than in mitochondria. On the whole, the results encourage the use of the Yoshida ascites hepatoma as a suitable model for studying protein turnover in relation to cell growth in vivo.

scholarly journals Versatile proteome labelling in fruit flies with SILAF

2019 ◽  
Author(s):  
Florian A. Schober ◽  
Ilian Atanassov ◽  
David Moore ◽  
Anna Wedell ◽  
Christoph Freyer ◽  
...  
Keyword(s):  
Protein Turnover ◽  
Cell Biology ◽  
Metabolic Regulation ◽  
Developmental Stages ◽  
Fruit Fly ◽  
Phosphorylation Sites ◽  
Turnover Rates ◽  
Post Translational Modification ◽  
Stable Isotope Labelling ◽  
Rapid Generation

ABSTRACTDrosophila melanogaster has been a working horse of genetics and cell biology for more than a century. However, proteomic-based methods have been limited due to technical obstacles, especially the lack of reliable labelling methods. Here, we advanced a chemically defined food source into stable-isotope labelling of amino acids in flies (SILAF). It allows for the rapid generation of a large number of flies with full incorporation of lysine-6. SILAF followed by fractionation and enrichment gave proteomic insights at a depth of 5,966 proteins and 7,496 phosphorylation sites, which substantiated metabolic regulation on enzymatic level. Furthermore, the label can be traced and predicts protein turnover rates during different developmental stages. The ease and versatility of the method actuates the fruit fly as an appealing model in proteomic and post-translational modification studies and it enlarges potential metabolic applications based on heavy amino acid diets.

scholarly journals Precise Estimation of In Vivo Protein Turnover Rates

2020 ◽  
Author(s):  
Jonathon J. O’Brien ◽  
Vikram Narayan ◽  
Yao Wong ◽  
Phillip Seitzer ◽  
Celeste M. Sandoval ◽  
...  
Keyword(s):  
Amino Acid ◽  
Protein Turnover ◽  
Search Algorithm ◽  
Deuterium Oxide ◽  
Isotopic Labeling ◽  
Analytical Framework ◽  
Turnover Rates ◽  
Ion Distribution ◽  
Common Technique

AbstractIsotopic labeling with deuterium oxide (D2O) is a common technique for estimating in vivo protein turnover, but its use has been limited by two long-standing problems: (1) identifying non-monoisotopic peptides; and (2) estimating protein turnover rates in the presence of dynamic amino acid enrichment. In this paper, we present a novel experimental and analytical framework for solving these two problems. Peptides with high probabilities of labeling in many amino acids present fragmentation spectra that frequently do not match the theoretical spectra used in standard identification algorithms. We resolve this difficulty using a modified search algorithm we call Conditional Ion Distribution Search (CIDS). Increased identifications from CIDS along with direct measurement of amino acid enrichment and statistical modeling that accounts for heterogeneous information across peptides, dramatically improves the accuracy and precision of half-life estimates. We benchmark the approach in cells, where near-complete labeling is possible, and conduct an in vivo experiment revealing, for the first time, differences in protein turnover between mice and naked mole-rats commensurate with their disparate longevity.

scholarly journals Interspecies differences in proteome turnover kinetics are correlated with lifespans and energetic demands

2020 ◽  
Author(s):  
Kyle Swovick ◽  
Denis Firsanov ◽  
Kevin A. Welle ◽  
Jennifer R. Hryhorenko ◽  
John P. Wise ◽  
...  
Keyword(s):  
Protein Turnover ◽  
Protein Misfolding ◽  
Bowhead Whale ◽  
Turnover Rates ◽  
Atp Production ◽  
Proteomic Approach ◽  
Mole Rat ◽  
Naked Mole Rat ◽  
Primary Fibroblasts ◽  
Energetic Demand

AbstractCells continually degrade and replace damaged and old proteins. However, the high energetic demand of protein turnover generates reactive oxygen species (ROS) that compromise the long-term health of the proteome. Thus, the relationship between aging, protein turnover and energetic demand remains unclear. Here, we used a proteomic approach to measure rates of protein turnover within primary fibroblasts isolated from a number of species with diverse lifespans including the longest-lives rodent, the naked mole rat and the longest-lived mammal, the bowhead whale. We show that organismal lifespan is negatively correlated with turnover rates of highly abundant proteins. In comparison to mice, cells from long-lived naked mole rats have slower rates of protein turnover, lower levels of ATP production and reduced ROS levels. Despite having slower rates of protein turnover, naked mole rat cells tolerate protein misfolding stress more effectively than mouse cells. We suggest that in lieu of rapid constitutive turnover, long-lived species may have evolved more energetically efficient mechanisms for selective detection and clearance of damaged proteins.

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