Regulation of branched-chain, and sulfur-containing amino acid metabolism by glutathione during ultradian metabolic oscillation of Saccharomyces cerevisiae.
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Autonomous ultradian metabolic oscillation (T approximately or =50 min) was detected in an aerobic chemostat culture of Saccharomyces cerevisiae. A pulse injection of GSH (a reduced form of glutathione) into the culture induced a perturbation in metabolic oscillation, with respiratory inhibition caused by H2S burst production. As the production of H2S in the culture was controlled by different amino acids, we attempted to characterize the effects of GSH on amino acid metabolism, particularly with regard to branched chain and sulfur-containing amino acids. During stable metabolic oscillation, concentrations of intracellular glutamate, aspartate, threonine, valine, leucine, isoleucine, and cysteine were observed to oscillate with the same periods of dissolved O2 oscillation, although the oscillation amplitudes and maximal phases were shown to differ. The methionine concentration was stably maintained at 0.05 mM. When GSH (100 microM) was injected into the culture, cellular levels of branched chain amino acids increased dramatically with continuous H2S production, whereas the cysteine and methionine concentrations were noticeably reduced. These results indicate that GSH-dependent perturbation occurs as the result of the promotion of branched chain amino acid synthesis and an attenuation of cysteine and methionine synthesis, both of which activate the generation of H2S. In a low sulfate medium containing 2.5 mM sulfate, the GSH injections did not result in perturbations of dissolved O2, NAD(P)H redox oscillations without burst H2 production. This suggests that GSH-dependent perturbation is intimately linked with the metabolism of branched-chain amino acids and H2 generation, rather than with direct GSH-GSSG redox control. (+info)
Estimation of the ideal ratio of true ileal digestible sulfur amino acids:lysine in 8- to 26-kg nursery pigs.
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Four experiments were conducted to determine the ideal ratio of true ileal digestible (TID) sulfur AA to Lys (SAA:LYS) in nursery pigs at two different BW ranges using both DL-Met and 2-hydroxy-4-(methylthio)-butanoic acid (HMTBA) as Met sources. In Exp. 1, 1,549 nursery pigs (Triumph 4 x PIC Camborough 22; initial BW 8.3 +/- 0.08 kg) were allotted to one of nine dietary treatments. The basal diet (Diet 1) was a semicomplex corn-soybean meal-based diet (1.32% TID Lys) with no supplemental HMTBA or DL-Met (47.7% TID SAA:LYS). Diets 2 to 9 consisted of the basal diet supplemented with four equimolar levels of DL-Met or HMTBA (52.7, 57.7, 62.7, and 67.7% TID SAA:LYS). In Exp. 2, 330 nursery pigs (Triumph 4 x PIC Camborough 22; initial BW 11.4 +/- 0.10 kg) were allotted to one of nine dietary treatments. The basal diet (Diet 1) was a corn-soybean meal-based diet (1.15% TID Lys) with no supplemental HMTBA or DL-Met (49% TID SAA:LYS). Diets 2 to 9 consisted of the basal diet supplemented with four equimolar levels of DL-Met or HMTBA (54, 59, 64, and 69% TID SAA:LYS). In Exp. 3, 1,544 nursery pigs (Triumph 4 x PIC Camborough 22; initial BW 12.4 +/- 0.13 kg) were allotted to one of nine dietary treatments as in Exp. 2. In Exp. 4, 343 nursery pigs (Genetiporc; initial BW 12.8 +/- 0.56 kg) were allotted to one of six dietary treatments. The basal diet (Diet 1) was a corn-soybean meal-based diet (1.05% TID Lys) with no supplemental DL-Met (49% TID SAA:LYS). Diets 2 to 5 consisted of the basal diet supplemented with four levels of DL-Met (54, 59, 64, and 69% TID SAA:LYS), and Diet 6 was the basal diet supplemented with one equimolar level of HMTBA to satisfy 59% TID SAA:LYS ratio. In all experiments, increasing the TID SAA:LYS ratio resulted in quadratic improvements in ADG (P < or = 0.09) and G:F (P < or = 0.05). Three different methods were used to estimate the optimal TID SAA:LYS ratio for each experiment. The two-slope broken-line regression model, x-intercept value of the broken-line and quadratic curve, and 95% of upper asymptote across the four experiments indicated that the average optimal TID SAA:LYS ratios were 59.3, 60.1, and 57.7% for ADG and 60.6, 61.7, and 60.1% for G:F, respectively. Thus, the optimal TID SAA:LYS ratio for 8- to 26-kg pigs based on the average value of these three estimates was 59.0% for ADG and 60.8% for G:F. (+info)
Effect of sulphur containing amino acids on [3H]-acetylcholine release from amacrine cells of the rabbit retina.
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1. The effects of the sulphur containing amino acids, homocysteic acid, homocysteine sulphinic acid, cysteic acid and cysteine sulphinic acid on the release of [3H]-acetylcholine ([3H]-ACh) from the cholinergic amacrine cells of the rabbit retina were examined. 2. All the compounds stimulated the spontaneous resting release and abolished the light-evoked release of [3H]-ACh. Except for homocysteine sulphinic acid these actions occurred at concentrations that did not affect the erg b-wave amplitude, indicating a site of action at the inner retina. 3. N-methyl-D-aspartate (in Mg(2+)-containing medium) clearly blocked the effects of homocysteic acid and homocysteine sulphinic acid on the resting release of [3H]-ACh but had no effect on the actions of cysteic acid and cysteine sulphinic acid. 4. Since N-methyl-D-aspartate is an antagonist of the light-evoked endogenous bipolar cell transmitter released onto cholinergic cells, these results are consistent with the suggestion that homocysteic acid or homocysteine sulphinic acid may be a transmitter released from this subpopulation of bipolar cells. 5. The present experiments indicate the existence of excitatory amino acids that have closer pharmacological properties to a bipolar cell transmitter than glutamate but it remains to be seen whether homocysteic acid or homocysteine sulphinic acid occur in these particular bipolar cells. (+info)
Comparative nutrition and metabolism: explication of open questions with emphasis on protein and amino acids.
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The 20th century saw numerous important discoveries in the nutritional sciences. Nonetheless, many unresolved questions still remain. Fifteen questions dealing with amino acid nutrition and metabolism are posed in this review. The first six deal with the functionality of sulfur amino acids (methionine and cysteine) and related compounds. Other unresolved problems that are discussed include priorities of use for amino acids having multiple functions; interactions among lysine, niacin and tryptophan; amino acid contributions to requirements from gut biosynthesis; the potential for gluconeogenesis to divert amino acids away from protein synthesis; the unique nutritional and metabolic idiosyncrasies of feline species, with emphasis on arginine; controversies surrounding human amino acid requirements; and the potential for maternal diet to influence sex ratio of offspring. (+info)
Methionine requirement of pigs between 5 and 20 kilograms body weight.
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Four nursery experiments were conducted using a methionine (Met)-deficient feather meal-corn-soybean meal-dried whey basal diet (20% CP; 3,250 kcal of ME/kg, .11% choline, .19% Met, 1.00% cystine) supplemented with lysine, tryptophan, and histidine to determine the Met requirement of 5- to 10- and 10- to 20-kg pigs. Based on a true Met digestibility value of 81.6% estimated by a pig ileal digestibility assay, the Met-deficient basal diet contained .155% of digestible Met. A preliminary experiment (Exp. 1) indicated that pigs fed the Met-deficient basal diet when fortified adequately with Met could produce weight gains similar to those of pigs fed a 20% CP practical corn-soybean meal-dried whey diet. In Exp. 2 and 3, crossbred pigs weighing 5.8 kg initially were fed diets containing graded levels of digestible Met between .195 and .355%. Average daily gain increased quadratically (P less than .05) as the level of Met increased. When the data of Exp. 2 and 3 were examined together, the digestible Met requirement of 5- to 10-kg pigs was estimated to be .255% of the diet. In Exp. 4 and 5, crossbred pigs averaging 10 kg were fed digestible Met concentrations ranging from .155 to .315%. Average daily gain increased quadratically (P less than .05). The digestible Met requirement of 10- to 20-kg pigs was estimated at .255% for maximal weight gain, which was similar to that of 5- to 10-kg pigs. Assuming an 89% digestibility of Met in practical corn-soybean meal diets, the total Met level needed in practice would be .29%.(ABSTRACT TRUNCATED AT 250 WORDS) (+info)
Rck2 is required for reprogramming of ribosomes during oxidative stress.
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Rck2 is a mitogen-activated protein kinase-activated protein kinase in yeast implicated in translational regulation. rck2Delta mutants are mildly sensitive to oxidative stress, a condition that causes dissociation of actively translating ribosomes (polysomes). In rck2Delta cells, polysomes are lost to an even higher degree than in the wild-type upon stress. Cells overexpressing the catalytically inactive rck2-kd allele are highly sensitive to oxidative stress. In such cells, dissociation of polysomes upon stress was instead greatly delayed. The protein synthesis rate decreased to a similar degree as in wild-type cells, however, indicating that in rck2-kd cells, the polysome complexes were inactive. Array analyses of total and polysome-associated mRNAs revealed major deregulation of the translational machinery in rck2 mutant cells. This involves transcripts for cytosolic ribosomal proteins and for processing and assembly of ribosomes. In rck2Delta cells, weakly transcribed mRNAs associate more avidly with polysomes than in wild-type cells, whereas the opposite holds true for rck2-kd cells. This is consistent with perturbed regulation of translation elongation, which is predicted to alter the ratio between mRNAs with and without strong entry sites at ribosomes. We infer that imbalances in the translational apparatus are a major reason for the inability of these cells to respond to stress. (+info)
Plant sulfite oxidase as novel producer of H2O2: combination of enzyme catalysis with a subsequent non-enzymatic reaction step.
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Sulfite oxidase (EC 1.8.3.1) from the plant Arabidopsis thaliana is the smallest eukaryotic molybdenum enzyme consisting of a molybdenum cofactor-binding domain but lacking the heme domain that is known from vertebrate sulfite oxidase. While vertebrate sulfite oxidase is a mitochondrial enzyme with cytochrome c as the physiological electron acceptor, plant sulfite oxidase is localized in peroxisomes and does not react with cytochrome c. Here we describe results that identified oxygen as the terminal electron acceptor for plant sulfite oxidase and hydrogen peroxide as the product of this reaction in addition to sulfate. The latter finding might explain the peroxisomal localization of plant sulfite oxidase. 18O labeling experiments and the use of catalase provided evidence that plant sulfite oxidase combines its catalytic reaction with a subsequent non-enzymatic step where its reaction product hydrogen peroxide oxidizes another molecule of sulfite. In vitro, for each catalytic cycle plant SO will bring about the oxidation of two molecules of sulfite by one molecule of oxygen. In the plant, sulfite oxidase could be responsible for removing sulfite as a toxic metabolite, which might represent a means to protect the cell against excess of sulfite derived from SO2 gas in the atmosphere (acid rain) or during the decomposition of sulfur-containing amino acids. Finally we present a model for the metabolic interaction between sulfite and catalase in the peroxisome. (+info)
Total sulfur amino acid requirement of healthy school-age children as determined by indicator amino acid oxidation technique.
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BACKGROUND: Current total sulfur amino acid (TSAA) requirements of children are based on a factorial estimate that involves several assumptions. OBJECTIVE: The objective was to determine the TSAA requirement (methionine alone) of healthy school-age children by measuring the appearance of 13CO2 (F13CO2) in breath after the oxidation of l-[1-13C]phenylalanine in response to graded methionine intakes. DESIGN: Six healthy school-age children randomly received each of 6 methionine intakes (0, 5, 10, 15, 25, and 35 mg.kg(-1).d(-1)) along with an amino acid mixture to give a final protein intake of 1.5 g.kg(-1).d(-1) and an energy intake of 1.7 x resting energy expenditure. The diet was devoid of cysteine. The mean TSAA requirement was determined by applying a biphase linear regression crossover analysis on F13CO2 data, which identified a breakpoint at minimal F13CO2 in response to graded methionine intakes. RESULTS: The mean and population-safe (upper 95% CI) intakes of TSAA (as methionine) were determined to be 12.9 and 17.2 mg.kg(-1).d(-1), respectively. CONCLUSIONS: The current study suggests that children of this age group have a mean TSAA requirement similar to that of adults (12.6 mg.kg(-1).d(-1)). Therefore, it is valid to use a factorial approach, which assumes that maintenance requirements in childhood are similar to adult requirements, to estimate TSAA requirements in school-age children. (+info)