• Title/Summary/Keyword: APO

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Interaction of Cytochrome c and $Mn^{2+}$ -Cytochrome c Peroxidase

  • Kim, Mun-kyoung;M. Kwon;Kim, K.;Sanghwa Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 1999.06a
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    • pp.44-44
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    • 1999
  • Yeast cytochrome c peroxidase (CcP) was cloned and overexpressed in E. coli, and purified by a Ni$^{2+}$-affinity column. HoloCcP was obtained by reconstituting apoCcP with Mn$^{3+}$-protoporphyrin IX (MnPP). Electron paramagnetic resonance (EPR) spectra of spin-labeled holoCcP showed a slightly more immobilized signal than spin-labeled apoCcP.(omitted)

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Electrophoretic Behaviors of α-Lactalbumin and β-Lactoglobulin Mixtures Caused by Heat Treatment

  • Lee, You-Ra;Hong, Youn-Ho
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1041-1045
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    • 2003
  • In order to study the reaction behaviors of bovine $\alpha$-lactalbumin ($\alpha$-La), $\beta$-lactoglobulin ($\beta$-Lg), and their mixtures during heat treatment, samples were analyzed using native-polyacrylamide gel electrophoresis (Native-PAGE), sodium dodecylsulfate (SDS)-PAGE, and two-dimensional (2-D)-PAGE. The electrophoresis demonstrated that the loss of native-$\alpha$-La increased as temperature increased, and that the loss of apo-$\alpha$-La was slightly higher than that of holo-$\alpha$-La. The tests also showed that during heat treatment, a mixture of $\alpha$-La and $\beta$-Lg was less stable than $\alpha$-La alone. As such, it was assumed that $\beta$-Lg induced holo-$\alpha$-La to be less stable than apo-$\alpha$-La during heat treatment. The reaction behavior of $\alpha$-La (holo-, apo-form) during heat treatment showed similar patterns in the 2-D-PAGE electropherogram, but the mixture of $\alpha$-La and $\beta$-Lg created new bands. In particular, the results showed a greater loss of native $\alpha$-La in the holo-$\alpha$-La and $\beta$-Lg mixture than in the apo-$\alpha$-La and $\beta$-Lg mixture. Thus, it can be concluded that the holo-$\alpha$-La and $\beta$-Lg mixture was more intensively affected by heat treatment than other samples, and that free sulphydryl groups took part in the heat-induced denaturation.

Meta-analysis on the Blood Lipids as Risk Factors of Coronary Heart Diseases in Koreans (한국인 관상동맥성 심질환의 위험요인으로서 혈청지질에 관한 메타분석)

  • Kim, Ki-Soon;Kim, Yang-Ok;Park, Jong;Park, Jong-Ku;Kim, Chun-Bae;Jee, Sun-Ha;Ryu, So-Yeon
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.4
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    • pp.491-498
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    • 1999
  • Objectives : To determine the relations between seven blood lipids such as total cholesterol(TC), triglyceride(TG), HDL-cholesterol(HDL), LDL-cholesterol(LDL), apolipoprotein A-1(Apo A1), apolipoprotein B(Apo B) and lipoprotein(a)(Lp(A)) and the coronary heart diseases(CHD), the quantitative techniques of meta-analysis were applied to studios of blood lipids and CHD in Koreans. Methods : We searched the Korean and the English literature published from 1980 to August, 1997 by manual search and bibliography review. Information on sample size, study design, participant characteristics(gender, age) and blood lipid levels were abstracted by reviewers using inclusion criteria. Estimates of the effect sizes of blood lipid levels on CHD in Koreans and corresponding 95% confidence intervals were calculated using random-effect models. Results : We identified 16 case-control studies to apply meta-analysis. The overall effect sizes for CHD were 20.3(95% CI: 14.23-20.22) in TC, 24.8(95% CI: 12.6-36.86) in TG, 15.16(95% CI: 3.99-26.33) in LDL, -3.48(95% CI: -5.79 - -1.17) in HDL, -9.78(95% CI: -16.98 - -2.58) in Apo-a1, 17.88(95% CI: 9.72-26.05) in Apo B and 18.95(95% CI: 17.88-20.02) in Lp(a). Conclusions : Our results suggested that seven blood lipids were significantly associated with CHD in Koreans. Well-designed and prospective studies between blood lipids and CMD in Koreans should be peformed.

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Intranasal Insulin for Alzheimer's Disease and Amnestic Mild Cognitive Impairment: Systematic Review and Meta-analysis (알츠하이머병 및 건망증 경도 인지장애의 인슐린 비강투여: 체계적 문헌 고찰 및 메타분석)

  • Alnajjar, Sarah;Jin, Hye Kyung;Kang, Ji Eun;Park, So Hyun;Rhie, Sandy Jeong
    • Korean Journal of Clinical Pharmacy
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    • v.27 no.3
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    • pp.161-170
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    • 2017
  • Background: There is recent evidence that insulin resistance is responsible for increasing the risk of developing cognitive dysfunction. To systematically review the influence of intranasal insulin treatment on the cognitive function in Alzheimer's disease patients. Methods: Randomized controlled trials comparing the cognitive effects of intranasal insulin therapy in Alzheimer's disease patients with controlled interventions were retrieved from Pubmed, Medline, Embase and Cochrane library. Meta-analysis was conducted on the cognitive measurements with a subgroup analysis by dose, gender and apolipoprotein E allele 4 (ApoE ${\varepsilon}4$) status. Results: Seven randomized controlled trials were eligible for inclusion. Intranasal insulin had a positive influence on the cognitive function as compared to placebo without a statistical significance (standardized mean difference; SMD = 0.109; 95% confidence interval; CI -0.04 to 0.26; P=0.14). In subgroup analysis, a 20 IU dose of intranasal insulin induced a significant improvement in cognitive function (SMD = 0.14; 95% CI 0.05 to 0.24; P=0.004), but 40 IU did not show this effect (SMD = -0.01; 95% CI -0.11 to 0.09; P=0.82). ApoE ${\varepsilon}4$ positive patients showed a significant decline in cognitive function as compared to ApoE ${\varepsilon}4$ positive patients in the control group (SMD = -0.213; 95% CI -0.38 to -0.04; P=0.015). Such an effect was not apparent in ApoE ${\varepsilon}4$ negative patients. Gender had no influence on the cognitive outcomes. Conclusion: The results indicate that intranasal insulin may have beneficial effect in improving the cognitive function in Alzheimer's disease patients.

LJ-1888, a selective antagonist for the A3 adenosine receptor, ameliorates the development of atherosclerosis and hypercholesterolemia in apolipoprotein E knock-out mice

  • Park, Jong-Gil;Jeong, Se-Jin;Yu, Jinha;Kim, Gyudong;Jeong, Lak Shin;Oh, Goo Taeg
    • BMB Reports
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    • v.51 no.10
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    • pp.520-525
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    • 2018
  • Cardiovascular diseases arising from atherosclerosis are the leading causes of mortality and morbidity worldwide. Lipid-lowering agents have been developed in order to treat hypercholesterolemia, a major risk factor for atherosclerosis. However, the prevalence of cardiovascular diseases is increasing, indicating a need to identify novel therapeutic targets and develop new treatment agents. Adenosine receptors (ARs) are emerging as therapeutic targets in asthma, rheumatoid arthritis, cancer, ischemia, and inflammatory diseases. This study assessed whether LJ-1888, a selective antagonist for $A_3$ AR, can inhibit the development of atherosclerosis in apolipoprotein E knock-out ($ApoE^{-/-}$) mice who are fed a western diet. Plaque formation was significantly lower in $ApoE^{-/-}$ mice administered LJ-1888 than in mice not administered LJ-1888, without any associated liver damage. LJ-1888 treatment of $ApoE^{-/-}$ mice prevented western diet-induced hypercholesterolemia by markedly reducing low-density lipoprotein cholesterol levels and significantly increasing high-density lipoprotein cholesterol concentrations. Reduced hypercholesterolemia in $ApoE^{-/-}$ mice administered LJ-1888 was associated with the enhanced expression of genes involved in bile acid biosynthesis. These findings indicate that LJ-1888, a selective antagonist for $A_3$ AR, may be a novel candidate for the treatment of atherosclerosis and hypercholesterolemia.

Effects of Jowiseungcheung-tang Extract on the Lipid Metabolism, Anti-oxidation and Inflammatory Reflex High Fat Diet Obese Rats (조위승청탕(調胃升淸湯) 추출액이 비만유도 쥐의 지질대사, 항산화계 및 염증반응계에 미치는 영향)

  • Oh, Sung-Won;Kim, Byoung-Woo
    • The Journal of Internal Korean Medicine
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    • v.34 no.1
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    • pp.1-13
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    • 2013
  • Objectives : The purpose of this study was to investigate the effects of Jowiseungcheung-tang (JWSCT) extract on the lipid metabolism, anti-oxidation and inflammatory reflex. Methods : Male Sprague-Dawley rats were fed a high fat diet for 8 weeks and were randomly divided into 4 groups (10 mice in each group): control group, 100 mg/kg JWSCT group, 200 mg/kg JWSCT group, 300 mg/kg JWSCT group. The control group was administered 100 mg/kg of water, but the other three groups were administered 100, 200, 300 mg/kg JWSCT extract for 4 weeks. After 4 weeks, we measured lipid level, thiobarbituric acid reactive substance (TBARS), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and cytokines in plasma and liver. The gene expression level and the ratio of apo-B and apo-E were then investigated by way of reverse transcription -polymerase chain reaction (RT-PCR). Results : In the JWSCT group, compared with the control, free fatty acid, triglyceride, total cholesterol, LDL-cholesterol, TBARS, IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ decreased significantly in plasma and liver. However HDL-cholesterol, IL-10, GSH-Px, SOD and CAT increased. In the JWSCT group, compared with the control, the gene expression level and the ratio of apo-A and apo-E decreased significantly in the RT-PCR analysis. Conclusions : The extract of JWSCT has anti-obesity, anti-inflammatory and anti-oxidant effects.

NICKEL INCORPORATION INTO Klebsiella aerogenes UREASE (Klebsiella aerogenes Urease로의 닉켈의 도입)

  • Lee, Mann-Hyung-
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.69-80
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    • 1994
  • Although ureases play important roles in microbial nitrogen metabolism and in the pathogenesis of several human diseases, little is known of the mechanism of metallocenter biosynthesis in this Ni-Containing enzyme. Klebsiella aerogenes urease apo-protein was purified from cells grown in the absence of Ni. The purified apo-enzyme showed the same native molecular weight, charge, and subunit stoichiometry as the holo-enzyme. Chemical modification studies were consistent with histidinyl ligation of Ni. Apo-enzyme could not be activated by simple addition of Ni ions suggesting a requirement for a cellular factor. Deletion analysis showed that four accessory genes (ureD, ureE, ureF, and ureG) are necessary for the functional incorporation of the urease metallocenter. Whereas the $\Delta$ureD, $\Delta$ureF, and $\Delta$ureG mutants are inactive and their ureases lack Ni, the $\Delta$ureE mutants retain partial activity and their ureases possess corresponding lower levels of Ni. UreE and UreG peptides were identified by SDS-polyacrylamide gel comparisons of mutant and wild type cells and by N-terminal sequencing. UreD and UreF peptides, which are synthesized at ve교 low levels, were identified by using in vitro transcription/translation methods. Cotransformation of E. coli cells with the complementing plasmids confirmed that ureD and ureF gene products act in trans. UreE was purified and characterized. immunogold electron microscopic studies were used to localize UreE to the cytoplasm. Equilibrium dialysis studies of purified UreE with $^{63}$ NiC1$_2$ showed that it binds ~6 Ni in a specific manner with a $K_{d}$ of 9.6 $\pm$1.3 $\mu$M. Results from spectroscopic studies demonstrated that Ni ions are ligated by 5 histidinyl residues and a sixth N or O atom, consistent with participation of the polyhistidine tail at the carboxyl termini of the dimeric UreE in Ni binding. With these results and other known features of the urease-related gene products, a model for urease metallocenter biosynthesis is proposed in which UreE binds Ni and acts as a Ni donor to the urease apo-protein while UreG binds ATP and couples its Hydrolysis to the Ni incorporation process.ouples its Hydrolysis to the Ni incorporation process.s.

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Effects of Bovine α-Lactalbumin Added with Oleic Acid and Microbial Transglutaminase on Cancer Cell Apoptosis (올레산과 미생물 유래 트랜스글루타미나제를 첨가한 우유 알파-락트알부민의 암세포사멸 효과)

  • Jeong, Ji-Eun;Hong, Youn-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.3
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    • pp.310-319
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    • 2012
  • In order to investigate the biological effects of conformational changes in the folding state of bovine ${\alpha}$-lactalbumin (${\alpha}$-La), the protein was prepared and classified as apo form, microbial transglutaminase (MTGase) added form, or bovine ${\alpha}$-La made lethal to tumor cell (BAMLET) form. Apo ${\alpha}$-La form showed weaker cancer cell inhibitory activity (apoptosis) than native ${\alpha}$-La, which suggests that the metal ion-like $Ca^{2+}$ had a positive effect, whereas BAMLET form showed strong cancer cell apoptotic activity. The BAMLET form seemed to be a molten globule structure that increased hydrophobicity. MTGase added to apo ${\alpha}$-La polymer showed similar anti-cancer activity as native ${\alpha}$-La, and it was well hydrolyzed by digestive enzymes. NMR results showed that BAMLET interacted with oleic acid and produced a complex.

Solution State Structure of P1, the Mimetic Peptide Derived from IgM Antigen Apo B-100 by NMR

  • Kim, Gilhoon;Lee, Hyuk;Oh, Hyewon;Won, Hoshik
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.3
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    • pp.95-101
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    • 2016
  • Apolipoprotein B-100 (Apo-B100) is a major component of low density lipoprotein (LDL). Apo B-100 protein has 4,536 amino acid sequence and these amino acids are classified into peptide groups A to G with subsequent 20 amino acids (P1-P302). The peptide groups were act as immunoglobulin (Ig) antigens which oxidized via malondialdehyde (MDA). The mimetic peptide P1 (EEEMLENVSLVCPKDAT RFK) out of D-group peptides carrying the highest value of IgG antigens were selected for structural studies that may provide antigen specificity. Circular Dichroism (CD) spectra were measured for peptide secondary structure in the range of 190-250 nm. Experimental results show that P1 exhibit partial of ${\beta}-sheet$ and random coil structure. Homonuclear (COSY, TOCSY, NOESY) 2D-NMR experiments were carried out for NMR signal assignments and structure determination for P1. On the basis of these completely assigned NMR spectra and distance data, distance geometry (DG) and Molecular dynamics (MD) were carried out to determine the structures of P1. The proposed structure was selected by comparisons between experimental NOE spectra and back calculated 2D NOE results from determined structure showing acceptable agreement. The total Root-Mean-Square-Deviation (RMSD) value of P1 obtained upon superposition of all atoms was in the range $0.33{\AA}$. The solution state P1 has mixed structure of ${\beta}-sheet$ (Glu[1] to Cys[12]) and random coil (Pro[13] to Lys[20]). These NMR results are well consistent with secondary structure from experimental results of circular dichroism. Structural studies based on NMR may contribute to the studies of atherosclerosis and observed conformational characteristics of apo B-100 in LDL using monoclonal antibodies.