• BMB Reports
• /
• v.36 no.4
• /
• pp.421-425
• /
• 2003

The serine/threonine protein kinase family is a large and diverse group of enzymes that are involved in the regulation of multiple cellular pathways. Elevated kinase activity has been implicated in many diseases and frequently targeted for the development of pharmacological inhibitors. Therefore, non-radioactive antibody-based kinase assays that allow high throughput screening of compound libraries have been developed. However, they require a generation of antibodies against the phosphorylated form of a specific substrate. We report here a time-resolved fluorescence assay platform that utilizes a commercially-available generic anti-phosphothreonine antibody and permits assaying kinases that are able to phosporylate threonin residues on protein substrates. Using this approach, we developed an assay for Cdc7/Dbf4 kinase activity, determined the $K_m$ for ATP, and identified rottlerin as a non-ATP competitive inhibitor of this enzyme.

• Korean Journal of Food Science and Technology
• /
• v.8 no.3
• /
• pp.129-135
• /
• 1976

The amino acid compositions of polished barley grain were investigated for 16 varieties by using amino acid autoanalyzer and simple correlation analysis between them and between protein and amino acid per protein was done. 1) Limiting amino acid is lysine, leucine and phenylalanine are high but threonine and tyrosine are low. Total essential amino acids is high. 2) Protein is significantly correlated negatively with lysine arginine, total basic amino acids (at p=0.01) and threonine, alanine, aspartic acid (at p=0.05) and positively with phenylalanine (at p=0.01) proline and cystine (at p=0.05). 3) Lysine is positively and significantly correlated with arginine and aspartic acid indicating that aspartic acid is probable precursor of lysine and that high yielding varieties or fertilization for high yielding decrease aspartic acid pool resulting low lysine. 4) Lysine content is positively correlated with dye binding capacity (at p=0.01). 5) Tryptophan is positively (at p=0.01) and significantly correlated with histidine, total basic amino acids and arginine. 6) In essential amino acids lysine, tryptophan, threonine and valine simultaneously increase or decrease while aromatic amino acids, sulfur contained amino acids, isoleucine and leucine do so together.

• Journal of the Korean Chemical Society
• /
• v.39 no.2
• /
• pp.123-126
• /
• 1995

• Asian-Australasian Journal of Animal Sciences
• /
• v.11 no.1
• /
• pp.1-7
• /
• 1998

A total of 120 pigs $(L\;{\times}\;LW\;{\times}\;D)$ averaged 14.16 kg of body weight were reared under six dietary treatments to evaluate the effects of amino acid supplementation on their performances. Treatments were 1) 18% CP diet (control); 2) 15% CP+0.28% Lys (B, 15L); 3) B+0.06% Met (15LM); 4) B+0.13% Thr (15LT) ; 5) B+0.06% Met+0.13% Thr (15LMT); 6) B+0.06% Met+0.13% Thr+0.05% Trp (15LMTT). Each treatment had 4 replicates with 5 pigs per replicate. The daily weight gains were statistically similar in 15LMT, 15LMTT and control groups. Threonine and methionine supplementation resulted in improved growth performance and nutrient digestibilities, while tryptophan supplementation had little beneficial effect. However, the best feed conversion was found in the control group. Dry. matter and CP digestibilities were improved in the 15LMT and 15LMTT groups. Gross energy, crude fat and phosphorus digestibilities were not affected by the treatment. Among the 15% CP groups, nitrogen digestibility showed the tendency to be increased as the more synthetic amino acids were added. Dry matter and nitrogen excretions were significantly reduced by feeding low protein, amino acid fortified diets, while phosphorus excretion was not influenced. Essential amino acids digestibility was higher in 15LT, 15LMT and 15LMTT groups and lower in the control. The 15LT group showed the best lysine digestibility, and methionine digestibilities were higher in all treated groups than control one. Threonine digestibilities higher in 15LT 15LMT, and 15LMTT than the control. The result shows that threonine and methionine should be added to the diet containing 15% CP diet, fortified with lysine to get same performance of 18% CP diet.

• Asian-Australasian Journal of Animal Sciences
• /
• v.14 no.8
• /
• pp.1157-1163
• /
• 2001

First-litter gilts were used to determine how different dietary treatments during gestation affect the reproductive performance of gilts and immunity development of their piglets. Twenty-two crossbred $Landrace{\times}Yorkshire$ gilts were randomly assigned to three dietary treatments, Following conception, the gilts were fed experimental diets until farrowing occurred. The diet for treatment I was low protein diet (8% CP), treatment 2 had an additional supplement of 0.14% threonine that was added to the low protein diet, and treatment 3 was a control diet containing 12% CP. During gestation, net body weight gain of sows in treatment group 2 was higher than in treatment group 1 (p=0,075), However, during lactation there was no difference between all treatments groups on body weight loss and their live piglets at birth. Although milk IgG between treatments did not differ, treatment groups 2 and 3 were slightly higher than treatment group I was, Plasma IgG concentrations in piglets were however equal within all treatment groups at birth and at 7 days of age, at 21 days of age, it was higher in treatment group I than it was in the other two groups (p<0.01), Threonine supplementation to a low protein diet during gestation slightly increases milk IgG of sows, It is beneficial for piglets to acquire more passive immunity, but a suppressive effect was also noted on the endogenous IgG synthesis in piglets. A gestation diet of 8% CP for gilts can stimulate immuno-system of her piglets.

• Molecules and Cells
• /
• v.26 no.1
• /
• pp.41-47
• /
• 2008

Mitogen-activated protein kinase (MAPK) signaling is a crucial component of eukaryotic cells; it plays an important role in responses to extracelluar stimuli and in the regulation of various cellular activities. The signaling cascade is evolutionarily conserved in the eukaryotic kingdom from yeast to human. In response to a variety of extracellular signals, MAPK activity is known to be regulated via phosphorylation of a conserved $T{\times}Y$ motif at the activation loop in which both threonine and tyrosine residues are phosphorylated by the upstream kinase. However, the mechanism by which both residues are phosphorylated continues to remain elusive. In the budding yeast, Saccharomyces cerevisiae, Fus3 MAPK is involved in the mating signaling pathway. In order to elucidate the functional mechanism of MAPK activation, we quantitatively profiled phosphorylation of the $T{\times}Y$ motif in Fus3 using mass spectrometry (MS). We used synthetic heavy stable isotope-labeled phosphopeptides and nonphosphopeptides corresponding to the proteolytic $T{\times}Y$ motif of Fus3 and accompanying data-dependent tandem MS to quantitatively monitor dynamic changes in the phosphorylation events of MAPK. Phosphospecific immunoblotting and the MS data suggested that the tyrosine residue is dynamically phosphorylated upon stimulation and that this leads to dual phosphorylation. In contrast, the magnitude of threonine phosphorylation did not change significantly. However, the absence of a threonine residue leads to hyperphosphorylation of the tyrosine residue in the unstimulated condition, suggesting that the threonine residue contributes to the control of signaling noise.

• Molecules and Cells
• /
• v.26 no.2
• /
• pp.200-205
• /
• 2008

The mitogen-activated protein kinase (MAPK) signaling pathway is activated in response to extracellular stimuli and regulates various activities in eukaryotic cells. Following exposure to stimuli, MAPK is known to be activated via dual phosphorylation at a conserved TxY motif in the activation loop; both threonine and tyrosine residues are phosphorylated by an upstream kinase. However, the mechanism underlying dual phosphorylation is not clearly understood. In the budding yeast Saccharomyces cerevisiae, the Hog1 MAPK mediates the high-osmolarity glycerol (HOG) signaling pathway. Tandem mass spectrometry and phosphospecific immunoblotting were performed to quantitatively monitor the dynamic changes occurring in the phosphorylation status of the TxY motif of Hog1 on exposure to osmotic stress. The results of our study suggest that the tyrosine residue is preferentially and dynamically phosphorylated following stimulation, and this in turn leads to the dual phosphorylation. The tyrosine residue was hyperphosphorylated in the absence of a threonine residue; this result suggests that the threonine residue is critical for the control of signaling noise and adaptation to osmotic stress.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.14 no.3
• /
• pp.301-304
• /
• 1985

Changes of free amino acids and free sugars in steamed and roasted green tea were determined after heat treatment at $110^{\circ}C$. Sixteen kinds of free amino acids and four kinds of free sugar were analyzed by HPLC. Free amino acids isolated were aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, histidine, lysine and arginine. Free sugars were sucrose, glucose, fructose, and raffinose. After the heat treatment, free amino acids and free sugars were decreased considerably.

• Korean Journal of Poultry Science
• /
• v.47 no.2
• /
• pp.107-114
• /
• 2020

This study was conducted to investigate the variation in nutrient composition of oilseed meals and to develop prediction equations for amino acid concentrations. Energy and nutrient contents were determined in a total of 1,380 feed ingredient samples including copra byproducts, corn distillers, dried grains with solubles, palm kernel byproducts, and soybean meal. The ingredient samples were imported to the Republic of Korea between 2006 and 2015. Data were analyzed using the MIXED procedure of SAS. The regression procedure of SAS was used to generate the prediction equation for the lysine concentration using the crude protein (CP) concentration as an independent variable. The concentrations of moisture, gross energy, CP, ether extract, crude fiber, ash, calcium, phosphorus, lysine, methionine, cysteine, and threonine in tested oilseed meals differed (P<0.05) depending on producing countries. The prediction equations for amino acid concentrations (% as-is basis) in the oilseed meals are: lysine = -1.08 + 0.080 × CP (root mean square error = 0.244, R² = 0.924, and P<0.001); threonine = -0.297 + 0.044 × CP (root mean square error = 0.099, R² = 0.958, and P<0.001). In conclusion, energy and nutrient compositions vary in the oilseed meals depending on the producing countries. Moreover, the crude protein concentration can be used as a suitable independent variable for estimating lysine and threonine concentrations in the oilseed meals.

• BMB Reports
• /
• v.31 no.4
• /
• pp.399-404
• /
• 1998

In order to gain further insight on the relationship between structure and function of glutathione S-transferase (GST), the six active-site mutants, R13T, K44T, Q51A, Q64A, S65A, and D98A, of human GST P1-1 were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. The active-site mutants showed marked differences in substrate specificity. The substitution of Gln51 with threonine resulted in a drastic decrease in the specific activities to <10% of the wild-type value. The substitution of Arg13 with threonine resulted in more decreased specific activity toward cumene hydroperoxide and in the $I_{50}$ values of S-(2,4-dinitrophenyl) glutathione and benanstatin A. These results suggest that the substitution of Arg13 with threonine changes the conformation of the active site to increase the affinity for the product or electrophilic substrate. Lys44 seems to be in the vicinity of the H-site of hGST P1-1 or may contribute to some extents to the electrophile binding.