• The Korean Journal of Pharmacology
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• 제27권2호
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• pp.135-144
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• 1991

Using T-lymphocytes obtained from rat peripheral blood, we found that the 44kD/pI6.8 protein was the major phosphoprotein of T-lymphocytes under basal condition, and that the 44kD/pI6.3 protein was a new phosphoprotein appeared in T-lymphocytes stimulated with ${\beta}-agonist$. The phosphorylation of the 44kD/pI6.3 protein was also induced by forskolin but inhibited by H-8 pretreatment. To clarify the character of the 44kD/pI6.3 protein, we used Con-A and kinase inhibitors, H-7 and W-7. Con-A stimulation induced phosphorylation of 44kD/pI 6.3 protein but that was inhibited by W-7 pretreatment. The phosphorytation of 44kD/pI6.3 protein was not induced by the PKC activator, PMA. Instead, the phosphorylation of 44kD/pI6.8 protein was reduced by H-7, a PKC inhibitor. From the above results,it can be concluded that the 44kD/pI6.3 protein can be a common substrate for A-kinase and CaM kinase. The two dimensional tryptic peptide mapping revealed that the 44kD/pI6.8 and 44kD/pI6.3 proteins are different.

• Journal of The Korean Society of Grassland and Forage Science
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• 제20권4호
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• pp.265-274
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• 2000

To investigate the effects of anion replacement on NO3- concentration in Italian ryegrass. Plants weregrown hydroponically to the full vegetative stage. NO3 supply(control) was replaced with SO1- (Tl), C1-(T2) and water (T3) to during 14 days. The determination of inorganic nutrient uptake and quantification ofprincipal metabolites (nitrate. protein and sugar) followed. Relatively high uptake of ~ a a'n d Ca" for controlplants, K+ and POJ- for T2 plants, and C 1 for TI plants was observed, respectively. Proteins in shoot andstubble were relatively higher in control and TI plants, which coupled N source. In root proteins largelydecreased (especially in T3 plants) during experimental period. Sugars in shoot of all four treatments tendedto decrease during the first 7 days and recovered afterward. Sugars in stubble also markedly decreased duringthe first 7 days, while those in root was much less varied during experimental period. After 14 days oftreatment, nitrate concentration in shoot of control plants was 13mgIg FW. Comparing to control: nitrate inshoot reduced by 27%, 46% and 50% in TI, T2 and T3 plants, respectively. Dry weight was slightlyincreased or not significantly changed in control, T1 and T2 plants, while a significant decrease(31.3% ofcontrol) occurred in T3 plants.(Key words : Italian ryegrass, Anion replacement, Ion uptake, Protein, Sugar, Nitrate)ptake, Protein, Sugar, Nitrate)

• Korean Journal of Veterinary Research
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• 제38권3호
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• pp.508-517
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• 1998

There is evidence that the sympathetic nervous system exerts a control on thyroid function via an adrenergic innervation of thyroid cells. Although it is clear that the inhibitory effects of catecholamines result from an activation of ${\alpha}_1$-adrenoceptors, the mechanisms involved in ${\alpha}_1$-stimulation are not fully understood. The effects of methoxamine and protein kinase C (PKC) activator on the release of thyroxine ($T_4$) from mouse thyroid were studied to clarify the role of PKC in the regulation of $T_4$ release in vitro. The glands were incubated in the medium, samples of the medium were assayed for $T_4$ by EIA kits. Methoxamine inhibited the TSH-stimulated $T_4$ release. This inhibition was reversed by prazosin, an ${\alpha}_1$-adrenergic antagonist. Futhermore, the inhibitory effect of methoxamine on the $T_4$ release stimulated by TSH was prevented by chloroethylclonidine, an ${\alpha}_{1b}$-adrenoceptor antagonist, but not by WB4101, an ${\alpha}_{1a}$-adrenoceptor antagonist. Also methoxamine inhibited the forskolin-, cAMP- or IBMX-stimulated $T_4$ release. These inhibition were reversed by PKC inhibitors, such as staurosporine and $H_7$. PMA, a PKC activator, completely inhibited the TSH-stimulated $T_4$ release, and its inhibition was reversed by staurosporine and $H_7$, but not by chelerythrine. R59022 (a diacylglycerol kinase inhibitor), like methoxamine, also inhibited the TSH-stimulated $T_4$ release, and its inhibition was also reversed by staurosporine. The present study suggests that methoxamine inhibition of $T_4$ release from mouse thyroid can be induced by activation of the ${\alpha}_{1b}$-adrenoceptors and that it is mediated through the ${\alpha}_1$-adrenoceptor-stimulated PKC formation.

• Journal of the Korea Academia-Industrial cooperation Society
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• 제14권11호
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• pp.5778-5784
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• 2013

In order to develop anti-wrinkle agent, we measured the anti-oxidative activity of gallic acid (GA) from Paeonia suffruticosa Andrews and investigated its cytotoxicity in HaCaT cells and then investigated its effect on tumor necrosis factor alpha (TNF-${\alpha}$)-induced matrix metalloproteinase-1 (MMP-1) mRNA, protein expressions and secretion in same cells. GA showed anti-oxidative activity with $IC_{50}$ of 30 ${\mu}g/mL$ and its activity was higher than that of butylated hydroxyanisol (BHA). GA showed weak cytotoxicity with high concentration (200 ${\mu}g/mL$) in HaCaT cells. MMP-1 mRNA, protein expression and secretion induced by tumor necrosis factor alpha (TNF-${\alpha}$) in HaCaT cells were significantly decreased by treatment of GA with dose-dependent manner(p<0.05). Therefore, our findings suggest that GA can be useful as an active ingredient for cosmeceuticals of anti-wrinkle effects.

• Korean Journal of Veterinary Research
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• 제39권6호
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• pp.1073-1080
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• 1999

Previous studies suggested that the inhibition of thyroxine ($T_4$) release by ${\alpha}_1$-adrenoceptor and muscarinic receptor stimulation results in activated protein kinase C (PKC) from mouse and guinea pig thyroids. In the present study, the effect of carbachol, methoxamine, phorbol myristate acetate (PMA), and R59022 on the release of $T_4$ from the mouse, rat, and guinea pig thyroids was compared to clarify the role of PKC in the regulation of the release of $T_4$. The thyroids were incubated in the medium containing the test agents, samples of the medium were assayed for $T_4$ by EIA kits. Forskolin, an adenylate cyclase activator, chlorophenylthio-cAMP sodium, a membrane permeable analog of cAMP, and isobutyl-methylxanthine, a phosphodiesterase inhibitor, like TSH (thyroid stimulating hormone), enhaced the release of $T_4$ from the mouse, rat, and guinea pig thyroids. Methoxamine, an ${\alpha}_1$-adrenoceptor agonist, inhibited the TSH-stimulated release of $T_4$ in mouse, but not rat and guinea pig thyroids. In contrast, carbachol, a muscarinic receptor agonist, inhibited the release of $T_4$ in guinea pig, but not mouse and rat thyroids. These inhibition were reversed by prazosin, an ${\alpha}_1$-adrenoceptor antagonist or atropine, a muscarinic antagonist or $M_1$- and $M_3$-muscarinic antagonists, in mouse or guinea pig thyroids. In addition, staurosporine, a PKC inhibitor, reversed methoxamine or carbachol inhibition of TSH stimulation. Furthermore, PMA, a PKC activator, and R59022, a diacylglycerol (DAG) kinase inhibitor, inhibited the TSH-stimulated release of $T_4$ in mouse, rat, and guinea pig thyroids. These inhibition were blocked by staurosporine. These findings suggest that the activation of receptor or DAG inhibits TSH-stimulated $T_4$ release through a PKC-dependent mechanism in thyroid gland.

• Food Quality and Culture
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• 제3권1호
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• pp.45-48
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• 2009

The investigation assessed the influence of Compositae plants consumption on the protein profile in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in Sprague-Dawley rats by injection of STZ (45 mg/kg body weight) into tail vein. The rats were randomly assigned to five groups: normal and STZ-control fed an AIN-93 diet, and groups whose diets were supplemented with 10% Compositae powder containing Artemisia iwayomogi (A. iwayomogi), Atractylodes lancea (A. lancea) or Taraxacum mongolicum (T. mongolicum). To observe the effects of Compositae plants in the animal model, the levels of protein in liver, kidney, lung, pancreas, and brain were determined after 4 weeks. The level of protein in kidney increased significantly in rats receiving the A. iwayomogi- and T. mongolicum-supplemented diet compared to the STZ-control group. The level of protein in lung was increased significantly in the A. iwayomogi-supplemented group. Blood glucose level correlated well with brain protein level but did not correlate with other protein levels. Also, blood glucose correlated inversely with kidney, lung and brain protein levels. It is suggested that supplementation with A. iwayomogi in diabetic rats leads elevates protein in kidney and lung.

• Journal of Life Science
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• 제28권2호
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• pp.170-175
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• 2018

Parkinson's disease (PD) is the most common movement disorder and the second most common neurodegenerative disease after Alzheimer's disease. Approximately 5~10% of PD patients are familial PD cases. Leucine-rich repeat kinase 2 (LRRK2) has been known to be a causal gene of PD when it is mutated. LRRK2 contains the functional kinase and GTPase domains as well as leucine-rich repeat (LRR) and WD40 domains that are known to play critical roles for protein-protein interaction, suggesting that LRRK2-interacting proteins are important regulators for PD pathogenesis. In an effort to identify proteins interacting with LRRK2, we carried out co-immunoprecipitation of LRRK2 antibody using extracts of NIH3T3 cells that express LRRK2 at a relatively high level. The mass spectrometry analysis of a precipitated band revealed that the co-precipitated band was methionyl-tRNA synthetase (MRS), an ancient enzyme that transfers methionin to its cognate tRNA. The interaction of MRS with LRRK2 was confirmed again by co-immunoprecipitation of endogenous proteins and GST pull-down assays. However, LRRK2 did not phosphorylate recombinant MRS protein in in vitro kinase assays, and over-expression of LRRK2 or MRS did not affect the stability of its partner protein. Our data indicate that LRRK2 interacts with but does not phosphorylate MRS, and the stability of each partner is not affected by the other.

• The Journal of Internal Korean Medicine
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• 제28권3호
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• pp.409-420
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• 2007

Objectives : The purpose of this study was to identify the anti-tumor effects of realgar on various cancer cells through molecular biologic and cellular biologic methods. Materials & Methods : We used 5 kinds of cancer cell lines:stomach cancer cell (AGS), glioma cells (T98G, A172, SNU-489) and prostate cancer cells (LNCaP). We injected the boiled extract of realgar. $50{\mu}$g/ml and $100{\mu}$g/ml to culture media (ml) for 24 hours. We examined the morphological changes under an inverted microscope and a fluorescence microscope. We measured the suppressive effect on viability of 5 kinds of cancer cells via XTT assay. We examined the effect on the revelation of PARP cleavage, Bcl-2 protein and Bax protein by western blot analysis. Results : The extract of realgar caused markedly morphological changes on AGS, T98G, SNU-489, and LNCaP. All of them showed withdrawn and floating appearance. The suppressive effect on viability of AGS, T98G, A172, SNU-489, and LNCaP showed that each test group had more suppressive effect on viability of AGS, T98G, A172, SNU-489, and LNCaP than the control group, which was statistically significantly (p<0.01). The extract of realgar did not induce PARP cleavage in AGS, T98G, A 172, SNU-489, or LNCaP. In the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in AGS, T98G, SNU-489, and LNCaP treated with realgar. The protein levels of Bcl-2 decreased and the protein levels of Bax did not change in A172 treated with realgar. Conclusions : This experiment showed that realgar has anti-tumor effect on stomach cancer cells (AGS), glioma cells (T98G, SNU-489L and prostate cancer cells (LNCaP)

• Journal of the Korean Magnetic Resonance Society
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• 제22권1호
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• pp.18-25
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• 2018

Replication protein A (RPA) is an essential single-stranded DNA binding protein in DNA processing. It is known that N terminal domain of RPA70 (RPA70N) recruits various protein partners including damage-response proteins such as p53, ATRIP, Rad9, and MRE11. Although the common binding residues of RPA70N were revealed, dynamic properties of the protein are not studied yet. In this study, we measured $^{15}N$ relaxation parameters ($T_1,\;T_2$ and heteronuclear NOE) of human RPA70N and analyzed them using model-free analysis. Our data showed that the two loops near the binding site experience fast time scale motion while the binding site does not. It suggests that the protein binding surface of RPA70N is mostly rigid for minimizing entropy cost of binding and the loops can experience conformational changes.

• Asian-Australasian Journal of Animal Sciences
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• 제13권5호
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• pp.659-665
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• 2000

A nitrogen (N) balance trial was conducted to examine the effect of N deprivation on subsequent N retention, blood urea nitrogen (BUN) and IGF-I levels and the ratio of IGF binding protein (IGFBP)-3 to IGFBP-l and -2. Pigs in treatment (T) 1 were given diet A (2.39% N) and those in T2 and T3 were given diet B (1.31% N) and excreta were collected (period 1 (P1)). Pigs in T1 continued to receive diet A while diets for T2 and T3 were changed to diets A and C (2.74% N), respectively. The excreta were collected for two more periods (P2 and P3). During P1, pigs in T2 and T3 retained 50% less N (p<0.001) than those in T1. However, pigs provided T2 (p<0.01) and T3 (p<0.05) retained more N than those assigned to T1 during P2. Pigs in T3 tended to retain more (p=0.10) N than those receiving T2 for the same period. The BUN values were lower (p<0.05) for pigs assigned to T2 and T3 than T1 during P1 and P2. Both IGF-I and IGFBP ratios of pigs assigned to T1 were higher (p<0.05) than those given T2 and T3 during P1 but no differences were found during P2 and P3.