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MPC 11 cell에 TPA 처리로 유도되는 DNA 결합 단백질의 특성
임계택,이정채 全南大學校 農業科學技術硏究所 1996 農業科學技術硏究 Vol.31 No.-
Vimentin의 전사인자(transcriptional factor)에 대한 기초자료로소 그 발현정도를 알아보기 위해 MPC11 cell에 Phorbol ester, 12-0-tetradecanoy phorbol 13-acetate (TPA)를 처리한 다음 DNA 결합 단백질을 세포질과 핵으로부터 분리하여 추출 한 후 gel mobility shift assay 방법으로서 그 성격을 연구했다. TPA를 처리한 MPC11 cell에서 처리 후 3시간에 단백질이 발현되었으며 그 단백질은 vimentin으로 추측된다. 한편 DNA 결합 단백질은 NF1과 TFN보다는 AP2에 대해 더 큰 결합력을 가지고 있는데 이는 AP2가 vimentin 발현과정에 있어 어떤 specific promotor element와 상호작용을 하는 것으로 여겨지며, TPA 의 처리에 의해서 vimentin의 발현이 촉진됨을 추측할 수 있었다.
이기섭,임계택 全南大學校 農業科學技術硏究所 1997 農業科學技術硏究 Vol.32 No.-
S에 대한 생체의 반응을 유도단백질로 표현하여 생리적인 급여 수준을 결정할 목적으로 mouse를 model로 실험한 결과는 다음과 같다. Na₂S·9H₂O를 1mM, 2mM, 3mM로 농도로 쥐에게 급여한 후 3h, 6h, 12h, 24h, 48h 간격으로 근육, 비장, 간, 뇌를 채취하여 유도단백질의 생성을 측정하였을때 근육은 1mM의 12h에서 약 27KDa를, 1mM의 12h, 24h, 48h과 2mM의 3h, 6h에서 약 15KDa의 유도단백질을 생성 하였다. 비장은 1mM의 6h와 3mM의 12h, 24h에서 약 50KDa의 유도단백질을 생성 하였으며 간에서는 3mM에서 6h, 12h, 24h에 약 18KDa의 유도단백질을 생성 하였으나 뇌의 경우에는 유도단백질을 관찰 하지 못하였다. 따라서 S와 Se는 근육에서 유사한 유도단백질을 보였으며 S에 있어서 근육과 비장에서 비교적 민감하게 반응 하였고 뇌의 경우에는 반응이 없음을 알 수 있었다. We measured the amount of the induction proteins to see the physiological level of S(sulfur). Two induction proteins in the muscle were detected by sulfur treatment. One of them is 27KDa at the concentration of 1mM/12hr, the other is 15KDa at 2mM/3hr. In the spleen, we observed just one induction protein, 50KDa, at 1 mM /6hr and 3mM/12hr respectively. 18KDa induction protein was expressed at 3mM/6hr in liver. In the brain, we didn't find any inducton protein . Generally, the induction proteins by S treatment were expressed early at high concentration, while the expression time of protein was slowly at low concentration, comparison with the high concentrations. The tendencies of the expression of induction protein by S treatment were similar to the case of Se. 6)
Kye Taek Lim,Kyung Sun Hea,Jae Han Shim 한국응용생명화학회 2002 Journal of Applied Biological Chemistry (J. Appl. Vol.45 No.4
The ethanol extract of Rhus verniciflua S. was subsequently isolated and fractioned into two portions using H_2O and 99% ethanol as elution buffers through silica gel column chromatography. To study the antioxidative effect of Rhus verniciflua S. extracts
Anti-oxidative Effects of Glycoprotein Isolated from Solanum nigrum L.
Kye-Taek Lim,Kyung-Sun Heo 한국식품영양과학회 2004 Journal of medicinal food Vol.7 No.3
Glycoprotein from Solanum nigrum L. (SNL glycoprotein) was isolated and tested for anti-oxidative effects on oxygen free radicals using a 1, 1-Diphenyl-2-picrylhydrazyl (DPPH) assay. The free radical scavenging activities of the SNL glycoprotein are optimal in acidic pH and up to 60℃. However, it has minimal activities in the presence of EDTA; although such activities are not dependent on M2+ ions (Ca2+, Mn2+, and Mg2+) in the presence of EDTA. Interestingly, when SNL glycoprotein was treated with deactivation agents (pronase E and NaIO4), the DPPH radical scavenging activity was decreased compared to the SNL glycoprotein treatment alone. The anti-oxidative effects of SNL glycoprotein on superoxide anion and hydroxyl radical under the optimal conditions revealed that SNL glycoprotein has remarkable scavenging effects on both radicals, but exhibited slightly higher scavenging effects on superoxide anion generated by enzymatic hypoxanthine (HX) / xanthine oxidase (XO) system than on hydroxyl radicals generated by the Fenton reaction. However, SNL glycoprotein was more effective against hydroxyl radials in cell cultures (NIH/3T3). Consequently, 20 g/ml SNL glycoprotein has a scavenging ability against superoxide anion corresponding to the ascorbic acid. On the other hand, its hydroxyl radial scavenging corresponds to 0.1 g/ml catalase. From these results, we suggest that SNL glycoprotein has potent anti-oxidative potential.
Kye-Taek Lim Korean Chemical Society 1991 Bulletin of the Korean Chemical Society Vol.12 No.3
The changed isotropic absolute Raman intensities of the phosphate residue in the complexes of positive charge oligopeptides, lys-lys, arg-arg, lys-aromat-lys, negative charge diethyl phosphoric acid (DEP) and polyriboadenylic acid{poly(rA)} were reported and discussed. Our measurements showed that the absolute intensities of phosphate stretch vibration in complexes were different according to the reaction partners. Due to the partial electrical charge and molecular structure of oligopeptides for the complex formation lysine can interact more strongly than arginine when the reaction partners have short chain and no steric hindrance. Owing to these reasons the intensity of phosphate stretching vibration is very sensitive according to the circumstance of reaction. From our results we could suggest that we can discriminate any one of the the lysine and arginine in the complicated biological molecule during interaction between nucleotides and proteins. The activity of reaction of two basical oligopeptides is not quite similar for complex formation in aqueous solution. The activity of dipeptides depends upon the structure of molecule and environment for complex formation. Aromatic ring contributes to electrostatic interaction in complexes. The amount of the absolute intensity for pure stacking interaction is smaller than electrostatic interaction in macromolecular complexes.
Lim, Kye-Taek,Lee, Jeong-Chae,Jung, Hee-Young,Jo, Sung-Kyun The Korean Society of Toxicology Korea Environment 2000 Toxicological Research Vol.16 No.2
The ethanol extract of Rhus verniciflua Stokes (RVS), the Korean Lacquer tree, was subsequentely isolated and fractioned into two portions using distilled water (SED) and 99% ethanol (SEE) as elution buffers through silica gel column (4x28 em, 22 $\AA$. 28~200 mesh). To know the antioxidative effect of the RVS extracts, primary hepatocytes were exposed to hydroxyl radical generated by 20 mU/$m\ell$ glucose oxidase with SED or SEE for 4 hr. The addition of 100$\mu\textrm{g}$/$m\ell$ SED in culture medium showed good protection from glucose oxidase (GO)-mediated cytotoxicity of hepatocytes, showing approximately equivalent to control. When the hepatocytes were incubated with 100 $\mu\textrm{g}$/$m\ell$ SED or SEE only for 4 hr. the activities of cell-associated superoxide dismutase (SOD) and catalase were elevated up to 1.22 fold and 1.4 fold, respectively, compared to control. Further increase, 1.88fold in SOD activity or 1.64fold in catalase activity, was also observed when the hepatocytes were incubated with 100 units/$m\ell$ of commercial SOD or catalase for 4 hr. Moreover. the glucose oxidase-mediated cytotoxicity in cultured hepatocytes was generally reduced upon addition of lysate obtained from SED or SEE-stimulated hepatocytes in a dose-dependent manner. From these results, we suggest that, in cultured hepatocytes, RVS ethanol extract can efficiently reduce cytotoxicity induced by glucose oxidase and may increase the activity of cell-associated SOD and/or catalase, thereby preventing and/or scavenging superoxides and hydroxyl radicals in this experiment.
Neuronal Cytotoxicity of Oxygen Radical in Newborn Mouse Forebrain Culture
Lim, Kye-Taek,Park, Seung-Taeck,Choi, Min-Kyu,Chung, Yeun-Tai Korean Society of ToxicologyKorea Environmental Mu 1995 Toxicological Research Vol.11 No.2
The cytotoxic effects of hydrogen peroxide and neuroprotective effects of a variety of agents were investigated in newborn mouse forebrain tissue culture. In our experiments, oxygen radical was generated enzymatically by glucose oxidase and the values were expressed as a percentage of number of living cells by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Cytotoxicity of oxygen radicals was prevented by catalase and (N, N, N', N', -tetrakis (2-pyridylmethyl) ethylenediamine (TPEN), but N-tetra-ot-butyl-phenylnitrone (PBN), and deferoxamine (DFX), failed to show protective effects against oxygen radicals. Antagonists of the N-methyl-D-aspartate (NMDA) receptor, D-2-amino-5-phosphonovaleric acid (APV), 7-chlorokynurenic acid (CKA), and MK801 (a non-competitive NMDA antagonist) were also not effective in blocking neurotoxicity induced by glucose oxidase generated oxygen radicals.