• 한국자기공명학회논문지
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• 제14권2호
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• pp.117-126
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• 2010

Hsp33 is a molecular chaperone achieving a holdase activity upon response to a dual stress by heat and oxidation. Despite several crystal structures available, the activation process is not clearly understood, because the structure inactive Hsp33 as its reduced, zinc-bound, monomeric form has not been solved yet. Thus, we initiated structural investigation of the reduced Hsp33 monomer by NMR. In this study, to overcome the high molecular weight (33 kDa), the protein was triply isotope-[$^{13}C$, $^{15}N$, $^2H$]-labeled and its inactive, monomeric state was ensured. 2D-[$^1H$, $^{15}N$]-TROSY and a series of triple resonance spectra could be successfully obtained on a high-field (900 MHz) NMR machine with a cryoprobe. However, under all of the different conditions tested, the number of resonances observed was significantly less than that expected from the amino acid sequence. Thus, a possible contribution of dynamic conformational exchange leading to a line broadening is suggested that might be important for activation process of Hsp33.

• Applied Microscopy
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• 제24권4호
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• pp.41-51
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• 1994

The topology of the enzyme has been investigated by biochemical studies including chemical labeling and cross linking. Thirteen subunits(polypeptides) of the cytochrome-c-oxidase have localistic characteristics of existing in the matrix side or cytoplasmic side in the mitochondria. In order to observe the distribution of the enzyme subunit on the mitochondria membrane, immunogold-labeling methods were employed. Antibody was obtained from the serum of immunized rabbit with enzyme subunit antigen which was obtained from cytochrome-c-oxidase of the beef heart muscle mitochondria. Beef heart muscle tissue as a tissue antigen was stained with immunized rabbit IgG and protein A gold complex. Electron microscopy has identified the existance of cytochrome-c-oxidase subunit $Mt_I,\;Mt_{II}\;and\;Mt_{III}$ on the membrane of cristae and outer chamber of mitochondria and the subunit $C_{IV}$ on the membrane of cristae and matrix of mitochondria. Particularly, the subunit $C_{IV}$ was also observed to exist in the sarcoplasm of muscle tissue.

• Molecules and Cells
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• 제42권4호
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• pp.356-362
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• 2019

The binding of MS2 bacteriophage coat protein (MCP) to MS2 binding site (MBS) RNA stem-loop sequences has been widely used to label mRNA for live-cell imaging at single-molecule resolution. However, concerns have been raised recently from studies with budding yeast showing aberrant mRNA metabolism following the MS2-GFP labeling. To investigate the degradation pattern of MS2-GFP-labeled mRNA in mammalian cells and tissues, we used Northern blot analysis of ${\beta}$-actin mRNA extracted from the Actb-MBS knock-in and $MBS{\times}MCP$ hybrid mouse models. In the immortalized mouse embryonic cell lines and various organ tissues derived from the mouse models, we found no noticeable accumulation of decay products of ${\beta}$-actin mRNA compared with the wild-type mice. Our results suggest that accumulation of MBS RNA decay fragments does not always happen depending on the mRNA species and the model organisms used.

• 한국작물학회지
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• 제55권4호
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• pp.357-364
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• 2010

Kjeldahl method used in many materials from various plant parts to determine protein contents, is laborious and time-consuming and utilizes hazardous chemicals. Near-infrared (NIR) reflectance spectroscopy, a rapid and environmentally benign technique, was investigated as a potential method for the prediction of protein content. Near-infrared reflectance spectra(1100-2400 nm) of coarse cereal grains(n=100 for each germplasm) were obtained using a dispersive spectrometer as both of grain itself and flour ground, and total protein contents determined according to Kjeldahl method. Using multivariate analysis, a modified partial least-squares model was developed for prediction of protein contents. The model had a multiple coefficient of determination of 0.99, 0.99, 0.99, 0.96 and 0.99 for foxtail millet, sorghum, millet, adzuki bean and mung bean germplasm, respectively. The model was tested with independent validation samples (n=10 for each germplasm). All samples were predicted with the coefficient of determination of 0.99, 0.99, 0.99, 0.91 and 0.99 for foxtail millet, sorghum, millet, adzuki bean and mung bean germplasm, respectively. The results indicate that NIR reflectance spectroscopy is an accurate and efficient tool for determining protein content of diverse coarse cereal germplasm for nutrition labeling of nutritional value. On the other hands appropriate condition of cereal material to predict protein using NIR was flour condition of grains.

• Journal of Chest Surgery
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• 제38권12호
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• pp.835-843
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• 2005

배경: 분자학적 표지자들과 식도암과의 관련성에 대해서 아직 뚜렷하게 밝혀진 바가 없다. 본 연구에서는 먼저 p53, Cyclin D1과 Ki67 지수 등 종양 표식자가 국내 식도암 환자에서 수술 후 얻어진 식도암 조직의 병기, 악성도, 임파선 전이 여부 등과 어떠한 관련성이 있는가를 알아보고 이를 이용해 향후 국내 식도암 환자에 대한 예후 분석 및 치료방침 결정 그리고 조기발견에 이용할 수 있는지 알아보았다. 또한 본 연구는 분자학적 표지자들의 임상적용의 기초를 마련하고자 하는 목적으로 시행하였다. 대상 및 방법: 1992년 3월부터 수술 후 조직절편이 보존된 124명의 환자를 대상으로 하였다. 조직절편의 파라핀을 제거한 뒤에 p53 변이, Cyclin D1의 과발현, Ki67 지수에 대한 면역조직화학 염색법을 시행하였으며 조직슬라이드상에서 세포분열지수를 구하였다. 이 결과와 임상적인 변수들과의 상관관계를 분석하였고, 환자의 생존 및 재발 결과와 비교하였다. 결과: 환자의 성별, 나이, 병기, 종양 크기, 타장기 전이 여부, 국소적인 재발 여부와 p53의 변이, Cyclin D1의 과발현, Ki67 지수, 세포분열 지수에 따른 통계적으로 유의한 차이는 없었다. Ki67 지수는 40 미만인 경우와 40 이상인 경우로 나누었을 경우 통계적으로 유의하게 생존기간에 있어서 차이가 나는 것으로 나타났으나(p=0.011) 다른 표지자들은 생존기간에 유의한 차이를 보이지 않았다. 결론: 높은 Ki67지수는 식도의 편평상피암의 예후에 나쁜 영향을 미치는 것으로 나타났으나 다른 표지자들은 영향을 미치지 않았다 이번의 연구를 통해 분자학적인 지표들이 임상적으로 가치 있는 인자가 될 수 있다고 판단되었다.

• BMB Reports
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• 제39권4호
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• pp.412-417
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• 2006

We examined the intracellular localization of NS5 protein of Dendrolimus punctatus cytoplasmic polyhedrosis virus (DpCPV) by expressing NS5-GFP fusion protein and proteins from deletion mutants of NS5 in baculovirus recombinant infected insect Spodoptera frugiperda (Sf-9) cells. It was found that the NS5 protein was present at the plasma membrane of the cells, and that the N-terminal portion of the protein played a key role in the localization. A transmembrane region was identified to be present in the N-terminal portion of the protein, and the detailed transmembrane domain (SQIHMVWVKSGLVFF, 57-71aa) of N-terminal portion of NS5 was further determined, which was accorded with the predicted results, these findings suggested that NS5 might have an important function in viral life cycle.

• The Korean Journal of Physiology and Pharmacology
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• 제13권4호
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• pp.281-285
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• 2009

Rotenone, a mitochondrial complex I inhibitor, can induce the pathological features of Parkinson's disease (PD). In the present study, naringin, a grapefruit flavonoid, inhibited rotenone-induced cell death in human neuroblastoma SH-SY5Y cells. We assessed cell death and apoptosis by measuring mitogen-activated protein kinase (MAPKs) and caspase (CASPs) activities and by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Naringin also blocked rotenone-induced phosphorylation of Jun NH2-terminal protein kinase (JNK) and P38, and prevented changes in B-cell CLL/lymphoma 2 (BCL2) and BCL2-associated X protein (BAX) expression levels. In addition, naringin reduced the enzyme activity of caspase 3 and cleavages of caspase 9, poly (ADP-ribose) polymerase (PARP), and caspase 3. These results suggest that naringin has a neuroprotective effect on rotenone-induced cell death in human neuroblastoma SH-SY5Y cells.

• 대한한의학회지
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• 제31권2호
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• pp.137-148
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• 2010

Objective: This study aimed to evaluate the protective effects of Gamipalmi-hwan (GPH) on elastase-induced lung cell injury. Materials and Methods: As an in vitro model of emphysema, the current study was performed to investigate potential activity of GPH in regulating injury responses of A549 human type II cell line mediated by elastase treatment. Results: GPH treatment increased the number of A549 cells which was reduced by elastase digestion. Elastin protein level, which was reduced by elastase treatment, was increased by GPH treatment. Labeling intensity with caspase 3 protein in elastase-treated cells was reduced by GPH treatment. Both Erk1/2 and Cdc2 protein levels, which were decreased by elastase treatment, were increased to a level similar to that of the normal cells. mRNA levels encoding IL-$1{\beta}$ and TNF-$\alpha$ were increased by elastase and then down-regulated by GPH. Conclusion: The present data suggest that A549 cells are subjected to inflammatory damage by elastase and can be recovered by GPH treatment. Further studies examining the protective activity of GPH in elastase-treated lung tissue would be useful for therapeutic strategies of emphysema treatment.

• Bulletin of the Korean Chemical Society
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• 제30권11호
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• pp.2647-2650
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• 2009

Cold shock proteins (Csps) are a subgroup of the cold-induced proteins on reduction of the growth temperature below the physiological temperature. They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays important role in cold adaptations for the psychrophilic microorganism. Recently, Cold shock protein from psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified. Three dimensional structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but structures of psychrophilic Csps were not studied yet. Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in this study. CpCsp was expressed in E. coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and $^{15}N$-labeled proteins with high purity over 90% was obtained. Further study will be carried out to investigate the tertiary structure and dynamics of CpCsp.

• 한국환경보건학회지
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• 제30권1호
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• pp.29-40
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• 2004

This study was performed to investigate and assess the composition of mineral and macronutrient contents in Korean cow′s milk.48 individual farm raw milk, 10 plant raw milk and 29 market milk were collected from June to August in 2003. The minerals such as calcium(Ca), potassium(K), magnesium(Mg), sodium(Na), zinc(Zn), iron(Fe) and phosphorus(P) were determined by using atomic absorption spectrometer(AAS). The macronutrients such as fat, protein and lactose were tested by using IR spectrometer. The obtained analytical results of minerals(mg/100 g) and rnaetronutrients (％) are as follows：1. In case of raw cow′s milk ； Ca 113.56, K 144.09, Mg 10.86, Na 42.53, Zn 0.42, Fe 0.030, p 113.32, fat 3.85, protein 3.08, lactose 4.80,2. In case of market cow′s milk ; Ca 103.04, K 142.46, Mg 10.27, Na 43.21, Zn 0.40, Fe 0.034. p 97.30, fat 3.78, protein 3.05, lactose 4.70,3. In case of fortified market cow′s milk ; Ca 165.40, K 145.79, Mg 10.57. Na 42.55, Zn 0.57, Fe 0.414, p 94.68, fat 3.74, protein 3.08, lactose 4.68,4. In case of processed market cow′s milk ; Ca 134.72, K 142.74, Mg 10.33, Na 45.07, Zn 0.50, Fe 0.650, p 92.48, fat 3.72, protein 3.07, lactose 4.74. According to the group of market milk(milk, fortified market row′s milk, processed market cow′s milk), the mean concentration of Ca and Fe were significantly higher in fortified and processed milk than milk(p<0.05). There were no significant differences in macronutrient(fat, protein, lactose) and mineral contents between pasteurized milk and UHT(ultra high temperature) treated milk($\alpha$=0.05). The labeled "Nutritional Facts" of market milk were satisfied with "Labeling Standards for Livestock Products of Korea". The measured mean concentrations of Ca, Fe, Zn were generally higher than lower limit of labeled value(above 80% of labeled value). The mean concentration of sodium was lower than upper limit of labeled value(below 120％ of labeled value).