• Journal of Microbiology
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• 제42권4호
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• pp.328-335
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• 2004

The human immunodeficiency virus type 1 (HIV-I) Tat protein transduction domain (PTD), which con­tains rich arginine and lysine residues, is responsible for the highly efficient transduction of protein through the plasma membrane. In addition, it can be secreted from infected cells and has the ability to enter neighboring cells. When the PTD of Tat is fused to proteins and exogenously added to cells, the fusion protein can cross plasma membranes. Recent reports indicate that the endogenously expressed Tat fusion protein can demonstrate biodistribution of several proteins. However, intercellular transport and protein transduction have not been observed in some studies. Therefore, this study exam­ined the intercellular transport and protein transduction of the Tat protein. The results showed no evi­dence of intercellular transport (biodistribution) in a cell culture. Instead, the Tat fusion peptides were found to have a significant effect on the transduction and intercellular localization properties. This sug­gests that the HIV-1 PTD passes through the plasma membrane in one direction.

• Applied Microscopy
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• 제31권2호
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• pp.199-206
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• 2001

완두 자엽세포에 대한 저장단백질 과립의 발달은 단백질 저장 액포의 가장자리에 축적된 단백질의 fragmentation에 의해서 이루어지는 것으로 알려져 왔다. 그러나 최근 이 외에도 terminal dilation, transformation, denovo development등의 독립적인 과정이 각각 다른 시기에 관찰되므로서 단백과립의 발달은 이러한 여러 과정이 모두 나타나는 복합과정임이 알려졌다. 이러한 과정과는 별도로 종자발달의 이른시기에 소포체 내강에 축적되는 저장단백질과 여기서 발달하는 단백과립에 대하여 규명하고자 발달중인 종자의 자엽으로부터 단일 세포를 얻어서 면역세포 화학적 반응을 실시하였다. 그 결과 종자발달의 이른 시기에는 legumin이, 중간시기 이후 에는 vicilin이 축적되므로서 단백질이 축적된 소포체가 단백과립으로 발달하는 것으로 나타났다 소포체 내강에 존재하는 단백질인 $\alpha-Tip$은 비교적 늦은시기에, toneplast membrane protein인 PPase는 이른시기에 각각 면역 세포화학적 반응이 관찰되었다.

• 한국잡초학회지
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• 제15권3호
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• pp.206-213
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• 1995

Glyphosate를 토마토의 동화산물 공급부위에 처리하였을 때, 제초제결합 단백질인 QB 단백질을 Escherichia coli 내에서 ${\alpha}$-galactosidase가 발현되기 위해 lac Z 유전자의 3' 말단에 cloning된 시금치 psb A 유전자에 의해 발현되는 hybrid 단백질에 대한 항체를 형성시킨 후 이것을 이용하여 immunoblotting을 실시하였다. G1yphosate는 thylakoid 막의 Photosystem II내에 있는 D1 단백질의 붕괴에 영향을 주었다. LHC II 복합체내의 D1 단백질의 기능 이상은 glyphosate 의 다면발현적 효과였다.

• Molecules and Cells
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• 제26권2호
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• pp.206-211
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• 2008

HI0381 of Haemophilus influenzae was investigated by circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy. HI0381 is a 153-residue peptidoglycan-associated outer membrane lipoprotein, and a part of the larger Tol/Pal network. Here, we report its backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments, and secondary structure predictions. About 97% of all of the $^1HN$, $^{15}N$, $^{13}CO$, $^{13}C{\alpha}$, and $^{13}C{\beta}$ resonances covering 131 non-proline residues of the 134 residue, mature protein, were clarified by sequential and specific assignments. CSI and TALOS analyses revealed that HI0381 contains five ${\alpha}$-helices and five ${\beta}$-strands. To characterize the structure of HI0381, the effects of pH and salt concentration were investigated by CD. In addition, the structural changes occurring when HI0381 was in a membranous environment were investigated by comparing its HSQC spectra and CD data in buffer and in DPC micelles; the results showed that helix ${\alpha}4$ and strand ${\beta}4$ became aligned with the membrane. We conclude that the conformation of HI0381 is affected by the membrane environment, implying that its folded state is directly related to its function.

• BMB Reports
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• 제30권1호
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• pp.73-79
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• 1997

This study was designed to investigate the effects of dietary garlic powder on cytochrome P450 enzymes and membrane stability in murine hepatocarcinogenesis initiated by diethylnitrosamine (DEN). Male Sprague-Dawley rats received a single intraperitoneal injection of DEN (200 mg/kg body wt) dissolved in saline. After 2 weeks on a basal diet, animals were fed diets containing 0. 0.5. 2.0. or 5.0% garlic powder for 6 weeks, and were subjected to two-thirds partial hepatectomy. The areas of placental glutathione S-transferase (GST-P) positive foci were inhibited in rats fed with garlic diets. GST-P is the most effective marker for DEN-initiated lesions. Hepatic microsomal lipid peroxidation was significantly decreased in rats fed with 2.0 and 5.0% garlic powder diets compared with that observed in the control animals and hepatic microsomal glucose 6-phosphatase (G6Pase) activity was found to increase significantly in rats fed 0.5 and 2.0% garlic powder diets. Thus as little as 0.5% garlic powder has a positive effect on the stability of hepatic microsomal membranes. p-Nitrophenol hydroxylase (PNPH) activity and the level of cytochrome P450 2E1 protein in the hepatic microsomes from rats fed diets containing 2.0 and 5.0% garlic powder were much lower than those of control microsomes. Rats fed 5.0% garlic powder diets exhibited the lowest P450 2E1 activity and protein levels among groups. Pentoxyresorufin O-dealkylase activity and immunoblot (cytochrome P450 2B1) analyses were not different between groups. However, the levels of cytochrome P450 1A1/2 protein in rats fed 0.5 and 2.0% garlic powder were significantly induced compared to controls. These results suggest that 2.0% garlic powder is effective in inhibiting the areas of GST-P positive foci, modulating certain isoforms of cytochrome P450 enzymes and stabilizing the hepatic microsomal membrane. Thus, the selective modification of cytochrome P450 enzymes and membrane stability by dietary garlic powder may influence areas of GST-P positive foci and chemoprevention of post-initiation of rat hepatocarcinogenesis.

• Bulletin of the Korean Chemical Society
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• 제28권12호
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• pp.2266-2270
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• 2007

Electron transfer of a redox protein at a bare gold electrode is too slow to observe the redox peaks. A novel Nafion-riboflavin functional membrane was constructed during this study and electron transfer of cytochrome c, superoxide dismutase, and hemoglobin were carried out on the functional membrane-modified gold electrode with good stability and repeatability. The immobilized protein-modified electrodes showed quasireversible electrochemical redox behaviors with formal potentials of 0.150, 0.175, and 0.202 V versus Ag/AgCl for the cytochrome c, superoxide dismutase and hemoglobin, respectively. Whole experiment was carried out in the 50 mM MOPS buffer solution with pH 6.0 at 25 oC. For the immobilized protein, the cathodic transfer coefficients were 0.67, 0.68 and 0.67 and electron transfer-rate constants were evaluated to be 2.25, 2.23 and 2.5 s?1, respectively. Hydrogen peroxide concentration was measured by the peroxidase activity of hemoglobin and our experiment revealed that the enzyme was fully functional while immobilized on the Nafion-riboflavin membrane.

• Journal of Microbiology and Biotechnology
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• 제22권4호
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• pp.470-478
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• 2012

Recent genome comparisons of E. coli B and K-12 strains have indicated that the makeup of the cell envelopes in these two strains is quite different. Therefore, we analyzed and compared the envelope proteomes of E. coli BL21(DE3) and MG1655. A total of 165 protein spots, including 62 nonredundant proteins, were unambiguously identified by two-dimensional gel electrophoresis and mass spectrometry. Of these, 43 proteins were conserved between the two strains, whereas 4 and 16 strain-specific proteins were identified only in E. coli BL21(DE3) and MG1655, respectively. Additionally, 24 proteins showed more than 2-fold differences in intensities between the B and K-12 strains. The reference envelope proteome maps showed that E. coli envelope mainly contained channel proteins and lipoproteins. Interesting proteomic observations between the two strains were as follows: (i) B produced more OmpF porin with a larger pore size than K-12, indicating an increase in the membrane permeability; (ii) B produced higher amounts of lipoproteins, which facilitates the assembly of outer membrane ${\beta}$-barrel proteins; and (iii) motility- (FliC) and chemotaxis-related proteins (CheA and CheW) were detected only in K-12, which showed that E. coli B is restricted with regard to migration under unfavorable conditions. These differences may influence the permeability and integrity of the cell envelope, showing that E. coli B may be more susceptible than K-12 to certain stress conditions. Thus, these findings suggest that E. coli K-12 and its derivatives will be more favorable strains in certain biotechnological applications, such as cell surface display or membrane engineering studies.

• 대한수의학회지
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• 제37권3호
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• pp.555-568
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• 1997

Fowl typhoid caused by Salmonella gallinarum has increased dramatically since 1992 and has caused a great economic losses in chicken industry by characterizing with high mortality. In these studies, we investigated the immunogenicity and protectivity in chickens which were immunized with outer membrane protein(OMP) extracted from isolates of S gallinarum against challenge with live microorganism. Outer membrane proteins were composed of various sizes of molecular weight including 14K, 22K, 31K, 36K, 40K and 55K and the most of them responded strongly against rabbit antisera in immunoblot analysis. The chickens vaccinated with OMP or vaccinated with whole-cell combined with OMP($200{\mu}g$/chickens) complex showed higher delayed type hypersensitivity(DTH) response than that of whole-cell vaccinated group. The protective rates of OMP or whole-cell combined with OMP complex group against challenge of S gallinarum were higher (above 75%) than those (45~50%) of whole-cell vaccinated group. All vaccines were safe and the body weight-gains of all vaccinated groups were not significantly different (p<0.05) from those of nonvaccinated control group. In vitro tests, OMP stimulated both the proliferation of lymphocytes and T-lymphocytes, and OMP-induced lymphocyte proliferation was higher in the cells of the immunized chickens with OMP than in those from the control chickens.

• Restorative Dentistry and Endodontics
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• 제24권2호
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• pp.286-298
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• 1999

The endodontic-periodontic combined lesions have been difficult to get correct diagnosis and predictable treatment. This study was to make the experimental endodontic-periodontic combined defects in dogs for the study of the periodontal regeneration and to evaluate the efficacy of the enamel matrix protein and e-PTFE membrane in the experimental endodontic-periodontic combined defects. 5 mongrel dogs were used. The pulp chambers were opened and the plaque was inserted into the chambers to induce the periapical lesions on the mandibular second, third and fourth premolars of the dogs. 1 month later, the root canal treatments were done with gutta perch a and ZOE sealer. On the day of surgery, the periapical defects were standardized by trephine bur. The buccal dehiscence defects were made by the dental bur and bone chisels. The apicoectomy with retrofilling was done. The prepared roots were randomly selected for test and control groups. In the experimental groups, the enamel matrix derivative and e-PTFE membrane were used. Nothing was placed on the control group. Fluroscent labelling was used to evaluate the bone formation. After 4 and 12 weeks, the dogs were sacrificed and undecalcified sections were prepared and stained with toluidine blue. Those histologic sections were examined by fluorescent microscopy and light microscopy. The results were as follows. 1. In the control group, new bone was formed in the periapical defects and scarcely in the buccal dehiscence defects. New cementum was not detected at 4 and 12 weeks. 2. In the experimental groups, new bone, new cementum and periodontal ligament were found in the periapical and buccal dehiscence defects. The relative amount and the quality of the new bone, new cementum and periodontal ligament tissue that had formed on the experimental groups were superior to those of the control group. 3. The current observation implicated that e-PTFE membrane and enamel matrix protein could be the effective tools for the guided tissue regeneration of the endo-perio combined defects.

• 공업화학
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• 제9권7호
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• pp.1090-1097
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• 1998

국부 마취제로 이용되는 lidocaine 화합물들이 마취 효과를 나타내는 과정을 알아보기 위하여 세포벽을 구성하는 지질 이중막의 모사 시스템으로 공기/물 계면에 형성된 지질 단분자 막을 이용하여 lidocaine 화합물들이 지질 단분자 막의 팽창에 미치는 영향을 연구하였다. Lidocaine이 신경 세포와 접하게 되면 세포벽을 구성하는 지질 이중막을 팽창시켜 이중막에 함침된 단백질을 압축하여 이를 통한 이온 통과가 차단되어 신경 전달이 마비된다는 가설과, 단백질 이온 통로에 존재하는 lidocaine receptor에 직접 흡착되어 이온 통로를 막는다는 가설이 구체적 실험 없이 제안되었다. 본 연구에서는 두 가설 타당성을 증명하고자 리피드 단분자 막팽창에 lidocaine이 미치는 영향을 조사하였다. 실험 결과 유용성인 lidocaine은 phosphatidyl choline, sphingomyelin, DS-PL95E, lipoid의 단분자 막을 수축시켰으며 phosphatidyl ethanolamine은 특정 조성 범위에서만 단분자 막을 팽창시켰다. 반면 수용성인 lidocaine-HCl 염은 실험에 사용된 모든 지질의 단분자 막을 팽창시켰다.