• BMB Reports
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• v.30 no.3
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• pp.222-228
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• 1997

Protein phosphatase 2C (PP2C) is one of the four major serine/threonine phosphatases which is dependent on $Mg^{2+}$ for its activity. PP2C was purified from rat liver cytosol and its characteristics were investigated. The substrate employed for routine assay was $[^{32}P]casein$ phosphorylated by PKA. The purification process involved DEAE chromatography, ammonium sulfate fractionation, phenyl sepharose chromatography, sephacryl 5-200 gel filtration, and histone agarose chromatography. The SDS-PAGE of PP2C showed one major single protein band at a position corresponding to a molecular mass of 43 kd and the purification fold was 637. The enzyme showed a pH optimum of 8 and $K_M$ value was $1.9\;{\mu}M$. However, when the substrate was changed to $[^{32}P]histone$, the pH optimum was shifted to 7 and $K_M$ value was $2.3\;{\mu}M.\;Mg^{2+}$ was essential to the enzyme activity and okadaic acid did not exert any inhibitory effect on the enzyme. To examine residue in the active site of PP2C effects of some protein-modifying reagents were tested.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.528-530
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• 2002

We have previously reported recombinant deletion mutant was constructed from cycloinulo-oligosaccharide fructanotransferase(CFTase) gene of Xanthmonas oryzae MGL21. Purification of the mutant protein from E. coli and reation condition for the production of inulo-oligosaccharide(ISO) were studied. The molecular mass of the CFTase deletion mutant protein was estimated to be 90kDa by SDS- Polyacrylamide gel electrophoresis. Optimum reaction pH and temperature were pH6.5 and $40^{\circ}C$, respectively. Under optimal reaction conditions, endo-inulinase activating enzyme was rapidly produced ISO from inulin. Components were DP(degree of polymerization) 3and 4 with trace amount of smaller oligosaccharides.

• The Korean Journal of Zoology
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• v.37 no.2
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• pp.232-239
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• 1994

RecA protein plans a central role in homologous recombination and DNA repair in Escherichia cofi (E. colD. The function 8nd structure of this protein are universal in prokarvotes and also conserved in eukaryotes such as yeast. The RecA-like protein with 74 lInDa in size has already been identified and purified from a fission yeast Schizosaccharomyces pombe (5. pommel (Lee, 19911. From this study it was revealed that the RecA-like protein of 5. pombe was highly inducible to various DNA damaging agents and inhibitors of nucleotide pool svnthesizins enzymes. The cellular level of the 5. pombe RecA-like protein wi,u markedly increased, upto 5- to 10-fold, by treatment with various DNA-damains agents including ultraviolet (UV) light, methyl methanesulfonate WS),4-nitroquinoline-1-oxide (4-NQO), and mitomycin-C (MMC), similar to E. cofi RecA protein. Interestingly, the protein level was also increased by inhibitors of nucleotide pool forming enzlwnes such as methotrexate (MTX) and hvdroxvurea (HU). The most effective doses for the inducibility of 4-NQO, MMS, W, MMC, MTX, and HU were 0.2 Ug/ml, 30 mM, 200 J/ma, 0.4 $\mus/ml,$ 1 Ug/ml, and 100 mM, respectively. The range of effective duration time for the inducibilitv of RecA-like protein was from 270 to 450 mins. These results suggest that the 5. pombe RecA-like protein also platys an imortant role in cellular responses to DNA damage as in E. coli system.

• Journal of Nutrition and Health
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• v.37 no.9
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• pp.763-770
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• 2004

This study was designed to compare the anti-carcinogenic effect of conjugated linoleic acid isomers on tumor incidence, cell proliferation and the levels of thromboxane (TX) B$_2$, prostaglandin (PG) E$_2$ and 1,2-diacylglycerol (DAG), and the related enzyme expression of cyclooxygenase (COX)-2 and protein kinase C (PKC) in colonic mucosa of 1,2-dimethy- lhydrazine (DMH) -treated rats. One hundred eight male Sprague Dawley rats were randomly divided into 3 groups depending on the types of CLA isomers, i.e. control group (no CLA contained), c9t11 group (cis-9, trans-11 CLA contained), and t10c12 group (trans-10, cis-12 CLA contained). The experimental diet was composed of protein at 20%, carbohydrate at 56.2%, and fat at 14.5% including 1.0% CLA isomers by weight. The experimental diet was fed for 30 weeks with the initiation of intramuscular injection of DMH, which was injected twice a week for 6 weeks to give total dose of 180 mg per kg body weight. Two CLA isomers (c9, t11, t10, c12) significantly reduced tumor incidence and cell proliferation by reducing the protein expression of COX-2 and PKC, and the level of TXB$_2$, PGE$_2$, and DAG in colonic mucosa. However, there was no significant difference in anti-carcinogenic effect between c9t11-CLA and t10c12-CLA.

• Biomolecules & Therapeutics
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• v.30 no.5
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• pp.447-454
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• 2022

Few studies have evaluated the role of autophagy in the development of oxaliplatin (OXT) resistance in colon cancer cells. In this study, we compared the role of autophagy between SNU-C5 colon cancer cells and OXT-resistant SNU-C5 (SNU-C5/OXTR) cells. At the same concentration of OXT, the cytotoxicity of OXT or apoptosis was significantly reduced in SNU-C5/OXTR cells compared with that in SNU-C5 cells. Compared with SNU-C5 cells, SNU-C5/OXTR cells exhibited low levels of autophagy. The expression level of important autophagy proteins, such as autophagy-related protein 5 (Atg5), beclin-1, Atg7, microtubule-associated proteins 1A/1B light chain 3B I (LC3-I), and LC3-II, was significantly lower in SNU-C5/OXTR cells than that in SNU-C5 cells. The expression level of the autophagy-essential protein p62 was also lower in SNU-C5/OXTR cells than in SNU-C5 cells. In SNU-C5/OXTR cells, the production of intracellular reactive oxygen species (ROS) was significantly higher than that in SNU-C5 cells, and treatment with the ROS scavenger N-acetylcysteine restored the reduced autophagy levels. Furthermore, the expression of antioxidant-related nuclear factor erythroid 2-related factor 2 transcription factor, heme oxygenase-1, and Cu/Zn superoxide dismutase were also significantly increased in SNU-C5/OXTR cells. These findings suggest that autophagy is significantly reduced in SNU-C5/OXTR cells compared with SNU-C5 cells, which may be related to the production of ROS in OXT-resistant cells.

• Korean Journal of Poultry Science
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• v.44 no.2
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• pp.67-73
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• 2017

The purpose of our study was to determine the effects of varying levels of energy, protein, and amino acids on the performances of laying hens. A total of 240 Hy-Line Brown laying hens at 36 weeks of age were used in this 4-week feeding trial. The hens were randomly allocated to five treatment diets, with eight replications of six hens in each replicate cage. The treatment diets were as follows: A- basal diet + 18% crude protein, metabolizable energy 2,800 kcal, total (methionine + cysteine) 0.65%; B- basal diet + 17% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.59%; C- basal diet + 16.5% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.59%; D- basal diet + 16.5% crude protein, metabolizable energy 2,700 kcal, total (methionine + cysteine) 0.54%; and E- basal diet + 16% crude protein, metabolizable energy 2,680 kcal, total (methionine + cysteine) 0.54%. The study results revealed that the hen-day egg production of hens that were fed with low-energy diets (B, C, and D) was comparable with that of hens fed with high-energy diet A, whereas average daily feed intake in hens fed treatment diet D and E was significantly higher (P<0.05) than that in hens fed treatment diet A. Overall, the eggshell thickness was unaffected by any of the treatment diets. Egg weight was comparable among the treatment diets, except for treatment diet E. Haugh unit improved with decreasing levels of dietary energy, protein, and methionine + cysteine in the diet. We can summarize that laying hens fed with low dietary energy and low crude protein treatment diets B, C, and D had satisfactory performance compared with those fed with high-energy treatment diet A. This indicates that there is the potential to reduce feed costs by formulating diets with lower energy and low protein levels.

• KSBB Journal
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• v.16 no.4
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• pp.426-429
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• 2001

To utilize sardine protein more effectively, fish meat paste products mixing sardine protein concentrate with pollack frozen meat paste at the ratio 0%, 15%, 20% and 25% were produced, and the change of firmness, sensory evaluation and the properties of amino and fatty acid were investigated. The quantity of sardine protein and it was almost gushed out around one hour at 100$\^{C}$. The firmness of the meat paste product was found as 0.54% and was better when the concentrated sardine protein was added at the ratio 15% and it was much higher than just that of pollack meat paste. In that case, total amino acid was the highest as 90.701 mg/g from the point of view of the amino acid composition. In terms of the fatty acid composition, unsaturated fatty acid of raw and boiled sardine was 61,8634% and 61.9384% each. We could find out that the high value of C$\_$20:5/ and C$\_$22:6/ of raw sardine was 7.2931% and 27.7843%, respectively.

• Journal of Life Science
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• v.7 no.3
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• pp.167-171
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• 1997

As an initial effort to elucidate RNA: protein binding in a way to regulate translation initiation and phosphorylation, a cDNA encoding a double-stranded RNA binding factor (RBFII)was isolated from Hela ZAPII cDNA library by affinity screening using [$\alpha$$^{-32}$P] UMP-labeled HIV Rev-responsive element(RRE) RNA. The nucleotide sequence of RBF (or TRBP) cDNA except the 5’end. At the 5’end, This common ORF was fused in-frame to N-terminal residues of Lac-Z through a unique 138 nt sequence encoding 46residues in the case of RBFII and a 63 nt sequence encoding 21 residuces in the case of RBFI. The context of ATG appearing first in the sequences suggests that both these cDNA inserts are incomplete at the 5’end.

• Biomolecules & Therapeutics
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• v.4 no.1
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• pp.97-102
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• 1996

To investigate whether human $\beta$$_2$-adrenergic receptor devoid of the C-terminal two transmembrane helices retain its ligand binding activity and specificity, 5'780-bp DNA fragment of the receptor gene which encodes amino acid 1-260 of human $\beta$$_2$-adrenergic receptor was subcloned into the bacterial fusion protein expression vector and expressed as a form of glutathione-S-transferase (GST) fusion protein in E. coli DH5$\alpha$. The receptor fusion protein was expressed as a membrane bound form which was verified by SDS-PAGE and Western blot. The fusion protein expressed in this study specifically bound $\beta$-adrenergic receptor ligand [$^3$H] Dihydroalprenolol. In saturation ligand binding assay, the $K_{d}$ value was 7.6 nM which was similar to that of intact $\beta$$_2$-adrenergic receptor in normal animal tissue ( $K_{d}$=1~2 nM) and the $B_{max}$ value was 266 fmol/mg membrane protein. In competition binding assay, the order of binding affinity of various adrenergic receptor agonists to the fusion protein was isoproterenol》epinephrine norepinephrine, which was similar to that of intact receptor in normal animal tissue. These results suggest that N-terminal five transmembrane helices of the $\beta$$_2$-adrenergic receptor be sufficient to determine the ligand binding activity and specificity, irrespective of the presence or absence of the C-terminal two transmembrane helices.s.s.s.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.5
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• pp.525-533
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• 1992

To improve the food quality of traditional Jeung-pyun(Korean fermenfed rice cake), effect of mixed ingredients on the quality of protein enriched product was studied. Changes were evaluated in chemical and textural properties of protein enriched Jeung-pyun altered by fermentation time, temperature, and steaming time. The maximum volume wxpansion was noted in dough mixed with rice flour(100 g), soy flour(25 g), sucrose(20 g), Tak-ju(50 ml) and water(10 ml), and then fermented at $35^{\circ}C$ for 3 hours. Soy protein isolate(SPI) enriched rice dough had a maximum expanded volume when 15g(w/w) of SPI, 20g(w/w) of sucrose, 60ml(v/w) of Tak-ju and 50ml(v/w) of water added into 100g rice flour, and then fermented at $30^{\circ}C$ for 2 hours. Steaming after fermentation made the smaller volume of bulk $45^{\circ}C$-50% of the initial volume be showed just after fermentation. Protein enriched Jeung-pyun prepared under the maximal volume endowing conditions showed the best protein quality(protein digestibility and protein efficiency ratio) and starch structure which was susceptible towards enzyme reaction. The improved starch quality of protein enriched Jeung-pyun could be confirmed by reducing power, gelatinization degree and amylose content. Jeung-pyun riched with soy flour at 20% level or with SPI at 5% showed the best overall quality by sensory and textural property but it had a problem in browning.