• Molecules and Cells
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• v.19 no.3
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• pp.398-401
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• 2005

The bacterial enzyme UDP-N-acetylglucosamine enolpyruvyl transferase catalyzes the first committed step of peptidoglycan biosynthesis, i.e., transfer of enolpyruvate from phosphoenolpyruvate to UDP-N-acetyl-glucosamine. We have overexpressed the enzyme from Haemophilus influenzae in Escherichia coli and crystallized it in the apo-form, as well as in a complex with UDP-N-acetylglucosamine and fosfomycin using ammonium sulfate as the precipitant. X-ray diffraction data from a crystal of the apo-form were collected to $2.8{\AA}$ resolution at 293 K. The crystal quality was improved by co-crystallization with UDP-N-acetylglucosamine and fosfomycin. X-ray data to $2.2{\AA}$ have been collected at 100 K from a flash-frozen crystal of the complex. The complex crystals belong to the orthorhombic space group I222 (or $I2_12_12_1$) with unit-cell parameters of a = 63.7, b = 124.5, and $c=126.3{\AA}$. Assuming a monomer of the recombinant enzyme in the crystallographic asymmetric unit, the calculated Matthews parameter ($V_M$) is $2.71{\AA}^3Da^{-1}$ and solvent content is 54.6%.

• BMB Reports
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• v.39 no.6
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• pp.717-721
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• 2006

In vitro analyses of type I signal peptidase activities require protein precursors as substrates. Usually, these pre-proteins are expressed in vitro and cleavage of the signal sequence is followed by SDS polyacrylamide gel electrophoresis coupled with autoradiography. Radioactive amino acids have to be incorporated in the expressed protein, since the amount of the in vitro expressed protein is usually very low and processing of the signal peptide cannot be followed by SDS polyacrylamide gel electrophoresis alone. Here we describe a rapid and simple method to express large amounts of a protein precursor in E. coli. We have analyzed the effect of ionophors as well as of azide on the accumulation of expressed protein precursors. Azide blocks the function of SecA and the ionophors dissipate the electrochemical gradient across the cytoplasmic membrane of E. coli. Addition of azide ions resulted in the formation of inclusion bodies, highly enriched with pre-apo-plastocyanine. Plastocyanine is a soluble copper protein, which can be found in the periplasmic space of cyanobacteria as well as in the thylakoid lumen of cyanobacteria and chloroplasts, and the pre-protein contains a cleavable signal sequence at its N-terminus. After purification of cyanobacterial pre-apo-plastocyanine, its signal sequence can be cleaved off by the E. coli signal peptidase, and protein processing was followed on Coomassie stained SDS polyacrylamide gels. We are optimistic that the presented method can be further developed and applied.

• BMB Reports
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• v.42 no.9
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• pp.605-610
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• 2009

Plasma high-density lipoprotein cholesterol (HDL-C) levels are inversely correlated with the risk of cardiovascular disease, and are known to increase with repetitive exercise. In the current study, HDL fractions from athletes' sera were isolated and compared as a function of the type of sport (runners [n = 10], throwers [n = 10], wrestlers [n = 10], and weight lifters [n = 8]), and as an age- and gender-matched reference group (n = 14). Among athletes, HDL from runners had the strongest antioxidant activity. Immunodetection showed that runners and wrestlers had the highest levels of apoA-I and lowest levels of apoA-II in their HDL. Electron microscopy also revealed that HDL2 of runners and wrestlers were the largest in size. In conclusion, although all athlete groups had significantly better serum lipid/lipoprotein profiles than the reference group, runners and wrestlers had the most desirable lipoprotein function and structure, including antioxidant activity, HDL-associated enzyme activities and increased particle size.

• BMB Reports
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• v.43 no.8
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• pp.535-540
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• 2010

Patients with hemorrhagic fever with renal syndrome (HFRS) often exhibit altered serum lipid and lipoprotein profile during the oliguric phase of the disease. Serum lipid and lipoprotein profiles were assessed during the oliguric and recovery phases in six male patients with HFRS. In the oliguric phase of HFRS, the apolipoprotein (apo) C-III content in high-density lipoproteins (HDL) was elevated, whereas the apoA-I content was lowered. The level of expression and activity of antioxidant enzymes were severely reduced during the oliguric phase, while the cholesteryl ester transfer protein activity and protein level were unchanged between the phases. In the oliguric phase, electromobility of $HDL_2$ and $HDL_3$ was faster than in the recovery phase. Low-density lipoprotein (LDL) particle size was smaller and the distribution was less homogeneous. Patients with HFRS in the oliguric phase had severely modified lipoproteins in composition and metabolism.

• Parasites, Hosts and Diseases
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• v.49 no.3
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• pp.331-335
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• 2011

A tick survey was conducted to determine the relative abundance and distribution of ticks associated with selected mammals in the Republic of Korea (ROK) during 2008-2009. A total of 918 ticks were collected from 76 mammals (6 families, 9 species) captured at 6 provinces and 3 Metropolitan Cities in ROK. Haemaphysalis longicornis (54.4%) was the most frequently collected tick, followed by Haemaphysalis flava (28.5%), Ixodes nipponensis (7.6%), Ixodes pomerantzevi (4.8%), Ixodes persulcatus (4.6%), and Haemaphysalis japonica (0.1%). Adults (57.0%) and nymphs (28.7%) of Ixodes and Haemaphysalis spp. were collected most frequently from medium or large mammals in this survey, while few larvae (14.3%) were collected. Hydropotes inermis was the most frequently captured mammal (52.6%), with a 16.4 tick index and 5 of 6 species of ticks collected during this survey. H. longicornis (69.7%) was the predominant tick collected from H. inermis, followed by H. flava (22.2%), I. persulcatus (6.1%), I. nipponensis (1.8%), and H. japonica (0.2%).

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.6
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• pp.1035-1042
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• 2002

This study was conducted to investigate the effect of sodium intake on serum lipids and glucose. 20 young adult women were fed the diets containing 290.5 mEq (high-Na diet) and 51.3 mEq (low-Na diets) Na for 6 days, respectively. Serum lipids, glucose, and other parameters after high-Na diet and low-Na diet were compared. The results would be summarized as follows. The mean age, body weight, height, and blood pressure of the subjects were 22.9$\pm$2.5 years,54.7+6.6 kg, 160.0$\pm$4.8 cm, 110.3$\pm$7.7/67.5$\pm$19.7 mmHg, respectively. Body weight, BMI, and diastolic blood pressure were significantly higher at the end of high-Na diet than of low-Na diet (p<0.001 p<0.001, p<0.05). However, there were not significantly different in height and systolic blood pressure between high- and low-Na diet. Serum cholesterol, LDL-cholesterol, and HDL-cholesterol were not significantly different with Na intakes. Serum triglyceride was significantly higher at the end of high-Na diet than of low- Na diet (p<0.05) Serum apo A-I was significantly decreased in low-Na diet, while apo B was increased (p<0.001, p<0.001). Thrombin time and prothrombin time, blood aggregation time were significantly faster following low-Na diet (p<0.001, p<0.05). There was not significantly different in serum glucose between high- and low-Na diet. However, serum insulin was significantly higher following low-Na diet (p<0.01). It is concluded that diastolic blood pressure, serum triglyceride, serum apo A-I, blood aggregation time were decreased in low-Na diet, while serum apo B and serum insulin were increased. These results suggest that Na-restricted diet affects not only blood Pressure but other biochemical parameters in blood. Therefore, for the patients who need restricted Na diet, it would be suggested that various biochemical changes should be carefully considered along with dietary Na manipulation.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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• pp.1368-1374
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• 2005

To investigate the genetic relationships among various cattle breeds, bovine mtDNA D-loop region was used in 411 animals of 18 cattle breeds, including 8 Asian Bos taurus, 7 European Bos taurus, 1 Asian Bos indicus, and 2 African Bos indicus. The size of amplified PCR products from mtDNA D-loop region was 964 bp and the products were digested by 15 different restriction enzymes. Two different band patterns were identified in eight restriction enzymes (BstXI, Hae III, Msp I, Apa I, Taq I, Alu I, BamH I, EcoN I) and the rest of restriction enzymes showed more than 3 different band patterns among which Apo I and MspR9 resulted in 7 different restriction patterns. The genotypes, number of haplotype, effective number of haplotype, and degree of heterozygosity were analyzed. Based on all the PCR-RFLP data, different haplotypes were constructed and analyzed for calculating genetic distances between these breeds using Nei's unbiased method and constructing a phylogenetic tree.

• Journal of the Korean Magnetic Resonance Society
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• v.20 no.3
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• pp.76-81
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• 2016

Copper is an elemental ion in living organisms. CopP from Helicobacter Pylori (HpCopP) is a copper(I)-binding protein and was suggested as regulator of copper metabolism in vivo. Previously, the metal binding property of HpCopP for Ag(I), Cu(I), and Cu(II) as well as the tertiary structure of HpCopP was shown. In this study, the dynamic profiles of HpCopP depending on metal binding were studied using ${^1H}-^{15}N$ steady-state NOE analysis. The heteroNOE experiment was performed for apo-CopP or metal-bound CopP. The obtained NOE values were analyzed and compared to figure out the effect of metals on the structural flexibility of HpCopP. As a result, Ag(I) and Cu(I) ions improved the rigidity of the structure while Cu(II) ion increased the flexibility of the structure, suggesting the oxidation of the CXXC motif decreases the structural stability of HpCopP.

• Molecules and Cells
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• v.38 no.12
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• pp.1096-1104
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• 2015

Particulate $matter_{2.5}$ ($PM_{2.5}$) is notorious for its strong toxic effects on the cardiovascular, skin, nervous, and reproduction systems. However, the molecular mechanism by which $PM_{2.5}$ aggravates disease progression is poorly understood, especially in a water-soluble state. In the current study, we investigated the putative physiological effects of aqueous $PM_{2.5}$ solution on lipoprotein metabolism. Collected $PM_{2.5}$ from Seoul, Korea was dissolved in water, and the water extract (final 3 and 30 ppm) was treated to human serum lipoproteins, macrophages, and dermal cells. $PM_{2.5}$ extract resulted in degradation and aggregation of high-density lipoprotein (HDL) as well as low-density lipoprotein (LDL); apoA-I in HDL aggregated and apo-B in LDL disappeared. $PM_{2.5}$ treatment (final 30 ppm) also induced cellular uptake of oxidized LDL (oxLDL) into macrophages, especially in the presence of fructose (final 50 mM). Uptake of oxLDL along with production of reactive oxygen species was accelerated by $PM_{2.5}$ solution in a dose-dependent manner. Further, $PM_{2.5}$ solution caused cellular senescence in human dermal fibroblast cells. Microinjection of $PM_{2.5}$ solution into zebrafish embryos induced severe mortality accompanied by impairment of skeletal development. In conclusion, water extract of $PM_{2.5}$ induced oxidative stress as a precursor to cardiovascular toxicity, skin cell senescence, and embryonic toxicity via aggregation and proteolytic degradation of serum lipoproteins.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.8
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• pp.1145-1152
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• 2012

This experiment was conducted with male chicks to investigate the influence of hormones and nutrients on the development of fatty liver syndrome (FLS) as well as the effects of dietary lipotropic factors on hepatic fat accumulation and lipogenic enzyme gene expression. A total of two-hundred sixteen 4-wk-old Hy-Line male chicks were divided into six groups and fed an experimental diet (T1, low-energy diet with low levels of lipotropic factors; T2, high-energy diet with low levels of lipotropic factors; T3 and T5, low-energy diet with high levels of lipotropic factors; T4 and T6, high-energy diet with high levels of lipotropic factors) for six weeks. The chicks in T5 and T6 groups were treated with intramuscular injections of estradiol benzoate for three days prior to biopsy and clinical analysis of FLS. Chicks treated with estrogen had significantly greater liver weights than untreated chicks. The abdominal fat contents were increased in chicks consuming high-energy diets as compared to those consuming low-energy diets. Treatment with estrogen significantly increased the concentrations of serum cholesterol, triacylglycerol and phospholipid (p<0.05). The hepatic triacylglycerol levels were tenfold higher in the estrogen treated chicks than in the untreated chicks. There were no significant differences in malondialdehyde levels between the treatment groups. Estrogen treatment dramatically increased the levels of fatty acid synthetase, acetyl-CoA carboxylase and ApoB mRNA. The results indicated that treatment with exogenous estrogen in growing male chicks induced hepatic fat accumulation, which might be partially due to increased lipogenic enzyme gene expression.