• Title/Summary/Keyword: APO

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Different Functional and Structural Characteristics between ApoA-I and ApoA-4 in Lipid-Free and Reconstituted HDL State: ApoA-4 Showed Less Anti-Atherogenic Activity

  • Yoo, Jeong-Ah;Lee, Eun-Young;Park, Ji Yoon;Lee, Seung-Taek;Ham, Sihyun;Cho, Kyung-Hyun
    • Molecules and Cells
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    • v.38 no.6
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    • pp.573-579
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    • 2015
  • Apolipoprotein A-I and A-IV are protein constituents of high-density lipoproteins although their functional difference in lipoprotein metabolism is still unclear. To compare anti-atherogenic properties between apoA-I and apoA-4, we characterized both proteins in lipid-free and lipidbound state. In lipid-free state, apoA4 showed two distinct bands, around 78 and $67{\AA}$ on native gel electrophoresis, while apoA-I showed scattered band pattern less than $71{\AA}$. In reconstituted HDL (rHDL) state, apoA-4 showed three major bands around $101{\AA}$ and $113{\AA}$, while apoA-I-rHDL showed almost single band around $98{\AA}$ size. Lipid-free apoA-I showed 2.9-fold higher phospholipid binding ability than apoA-4. In lipid-free state, $BS_3$-crosslinking revealed that apoA-4 showed less multimerization tendency upto dimer, while apoA-I showed pentamerization. In rHDL state (95:1), apoA-4 was existed as dimer as like as apoA-I. With higher phospholipid content (255:1), five apoA-I and three apoA-4 were required to the bigger rHDL formation. Regardless of particle size, apoA-I-rHDL showed superior LCAT activation ability than apoA-4-rHDL. Uptake of acetylated LDL was inhibited by apoA-I in both lipid-free and lipid-bound state, while apoA-4 inhibited it only lipid-free state. ApoA-4 showed less anti-atherogenic activity with more sensitivity to glycation. In conclusion, apoA-4 showed inferior physiological functions in lipid-bound state, compared with those of apoA-I, to induce more pro-atherosclerotic properties.

Synthesis of Reconstituted High Density Lipoprotein (rHDL) Containing apoA-I and apoC-III: the Functional Role of apoC-III in rHDL

  • Cho, Kyung-Hyun
    • Molecules and Cells
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    • v.27 no.3
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    • pp.291-297
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    • 2009
  • Apolipoprotein (apo) C-III is a marker protein of triacylglycerol (TG)-rich lipoproteins and high-density lipoproteins (HDL), and has been proposed as a risk factor of coronary heart disease. To compare the physiologic role of reconstituted HDL (rHDL) with or without apoC-III, we synthesized rHDL with molar ratios of apoA-I:apoC-III of 1:0, 1:0.5, 1:1, and 1:2. Increasing the apoC-III content in rHDL produced smaller rHDL particles with a lower number of apoA-I molecules. Furthermore, increasing the molar ratio of apoC-III in rHDL enhanced the surfactant-like properties and the ability to lyse dimyristoyl phosphatidylcholine. Furthermore, rHDL containing apoC-III was found to be more resistant to particle rearrangement in the presence of low-density lipoprotein (LDL) than rHDL that contained apoA-I alone. In addition, the lecithin:cholesterol acyltransferase (LCAT) activation ability was reduced as the apoC-III content of the rHDL increased; however, the CE transfer ability was not decreased by the increase of apoC-III. Finally, rHDL containing apoC-III aggravated the production of MDA in cell culture media, which led to increased cellular uptake of LDL. Thus, the addition of apoC-III to rHDL induced changes in the structural and functional properties of the rHDL, especially in particle size and rearrangement and LCAT activation. These alterations may lead to beneficial functions of HDL, which is involved in anti-atherogenic properties in the circulation.

Association of apolipoprotein E polymorphisms with serum lipid profiles in obese adolescent (비만아에서 고지혈증과 Apolipoprotein E 다형성의 관계)

  • Yoon, Jung Min;Lim, Jae Woo;Cheon, Eun Jung;Ko, Kyoung Og
    • Clinical and Experimental Pediatrics
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    • v.51 no.1
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    • pp.42-46
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    • 2008
  • Purpose : Apolipoprotein E (Apo E) plays a major role in lipoprotein metabolism and lipid transport. Many investigators have described that Apo E polymorphisms is one of the most important genetic determinants for cardiovascular disease. The purpose of this study was to evaluate the association between Apo E polymorphisms and serum lipid profiles in obese adolescent. Methods : We measured the serum concentrations of glucose, apolipoprotein (Apo) A1, Apo B, total cholesterol (TC), triglyceride (TG), HDL and LDL-cholesterol after overnight fasting in obese adolescent. Apo E polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results : 86 obese adolescents participated in this study. The body mass index (BMI) of participants were excess of 95 percentile by age and sex. Male to female ratio was 1.7 and mean age of study group was $16.2{\pm}1.8\;years$. Mean BMI was $27.4{\pm}2.5kg/m^2$. The frequency of ${\varepsilon}2$, ${\varepsilon}3$ and ${\varepsilon}4$ allele were 8.1%, 87.2% and 4.7% respectively. Study populations were classified into the following three genotypes 1) Apo E2 group (n=13, 15.1%) carrying either the ${\varepsilon}2/{\varepsilon}2$ or ${\varepsilon}2/{\varepsilon}3$ 2) Apo E3 group (n=65, 75.6%) carrying the most frequent ${\varepsilon}3/{\varepsilon}3$ 3) Apo E4 group (n=8, 9.3%) carrying either the ${\varepsilon}3/{\varepsilon}4$ or ${\varepsilon}4/{\varepsilon}4$. No differences were found among Apo E genotypes concerning age, sex, weight, height and BMI. Apo B and LDL-cholesterol concentrations were significantly higher in the Apo E4 group (P<0.05). No association were found between Apo E genotypes and glucose, Apo A1, TC, TG and HDL. Conclusions : We confirmed that serum concentrations Apo B and LDL-cholesterol were influenced by Apo E genotypes. Apo E polymorphisms seems to influence some alteration of lipid metabolism associated with obesity in adolescent.

ApoE Allele Test in Korean with Hair Root DNA (모근 DNA를 이용한 한국인의 ApoE 유전자형 검사)

  • Kim, Chong-Ho;Jung, Mi-Ra;Park, Sang-Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.179-183
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    • 2006
  • Alzheimer's disease (AD) is characterised neuropathologically by the accumulation of neuritic plaques and neurofibrillary tangles as well as by cerebrovascular amyloid deposition and neuronal cell loss. Current advances have shown the apolipoproteinE-epsilon 4 (ApoE4) allele to be highly associated with late-onset familial and sporadic Alzheimer's disease (AD) in Western populations. The association of ApoE allele frequencies and dementia remains unknown in populations from many countries. We recently initiated a project to examine ApoE frequencies in non-demented healthy Koreans. Genomic DNA in hair root from a thousand persons was collected and ApoE gene type was investigated with the methods of polymerase chain reaction (PCR) and restriction fragment length polymorphism. A group of a thousand non-demented Koreans over the age of 40 years were found to be positive in 15.7% of the cases for ApoE4. AD and ApoE4 were closely related. ApoE epsilon 4 was a dangerous factor of AD and ApoE 4 allele made a contribution to the heterogenicity of AD.

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Apolipophorin-III uptake by the last larval fat body in the wax moth Galleria mellonella (꿀벌부채명나방 종령 유충 지방체에 의한 아포리포포린-III의 흡수)

  • Yun, Hwa-Kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.8
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    • pp.4106-4110
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    • 2013
  • Apolipophorin-III (apoLp-III) was isolated and purified from the last larval hemolymph of Galleria mellonella by the KBr gradient ultracentrifugation and gel chromatography (Sephadex G-100). In this paper, we examined that apoLp-III is taken up into the last larval fat bodies in Galleria mellonella. The last larval fat body tissues were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III (FITC-apoLp-III). Fluorescein microscopy and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that the last larval fat body tissues internalize FITC-apoLp-III. The results show that the apoLp-III is taken up by the last larval fat body.

Formation of Oxidative Cleavage Products from Lycopene (Lycopene으로부터 산화개열산물의 생성)

  • Kim, Seon-Jae
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1227-1233
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    • 2000
  • Lycopene was subjected to ozonolysis in ice-cold dichloromethane. The ozonolysis products were fractionated with a silica column and the carbonyl fraction was analyzed by ODS-HPLC with a photodiode array detector and by LC-MS. UV-vis spectra and $[M+H]^+$ of the carbonyl compound peaks showed clearly that acycloretinal, apo-14'-lycopenal, apo-12'-lycopenal, apo-10'-lycopenal, apo-8'-lycopenal and apo-6'-lycopenal were formed by ozonolysis of lycopene. Lycopene was solubilized in toluene and aqueous Tween 40, and then oxidized by incubating at $37^{\circ}C$ under atmospheric oxygen. Carbonyl compounds were produced. In comparison with autoxidation and ozonolysis, each compound showed the same retention time and UV-vis spectra are identical to the reference cleavage products prepared by ozonolysis of lycopene. Thus, eccentric cleavage of lycopene was confirmed to occur in vitro under oxidation condition.

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Effects of Apolipoprotein A-I on Apoptosis and Cytokine Production in Human Neutrophils (인간 호중구의 세포사멸과 시토카인 분비에 대한 아포지방단백 A-I의 영향)

  • Kang, Hyung-Kon;Choi, Jae-Hyung;Huh, Jae-Taeck
    • Journal of Life Science
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    • v.21 no.1
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    • pp.22-30
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    • 2011
  • Apolipoprotein A-I (apoA-I) has anti-inflammatory and anti-oxidative properties. This study was designed to investigate whether apoA-I affects apoptosis and cytokine production of human blood neutrophils in an in vitro culture system. Spontaneous apoptosis of neutrophils was significantly delayed by apoA-I. In addition, high density lipoprotein containing apoA-I also delayed apoptosis of neutrophils. Apoptosis of neutrophils was inhibited by anti-scavenger receptor type B-I antibodies. The amounts of interleukin-8, interferon (IFN)-inducible protein 10 (IP-10), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$) in the supernatants of cultured neutrophils treated with apoA-I were significantly increased. Combined treatment of neutrophils with IFN-$\gamma$ and apoA-I produced higher amounts of IP-10 and TNF-$\alpha$ than did treatment with IFN-$\gamma$ or apoA-I alone. The present study reveals that apoA-I activates neutrophils to produce cytokines and delays spontaneous apoptosis of neutrophils. These findings suggest that apoA-I, although a well-known negative acute-phase protein, has a pro-inflammatory effect in neutrophils.

Apolipophorin-III uptake by the adult testes in the wax moth, Galleria mellonella (꿀벌부채명나방 성충 정소에 의한 아포리포포린-III의 흡수)

  • Yun, Hwa-Kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.10
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    • pp.199-203
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    • 2016
  • Apolipophorin-III (apoLp-III) was isolated and purified from the last larval hemolymph of Galleria mellonella by KBr gradient ultracentrifugation and gel chromatography (Sephadex G-100). After KBr gradient ultracentrifugation, the lipophorin-free fractions were used as the samples for gel chromatography. The purity of the finally purified apoLp-III was confirmed by SDS-PAGE after gel chromatography. In this study, we found that apoLp-III is taken up into the adult testes in Galleria mellonella. The testes were dissected from day-1 or -2 adults in cold Ringer's solution and used for tissue culture. The protein moiety of apoLp-III was labeled with FITC dissolved in dimethyl sulfoxide (DMSO) at room temperature under conditions of continuous stirring for 1 h. The FITC-labeled apoLp-III was purified with a Sephadex G-25 PD-10 column. The tissues of the adult testes were incubated at room temperature for 30 min with fluorescein isothiocyanate (FITC)-labeled apoLp-III. Fluorescein microscopy and sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) revealed that the adult testis tissues internalize the FITC-labeled apoLp-III. The results showed that apoLp-III is taken up by the adult testes.

Relationship Among Apolopoprotein E Phenotypes, Dietary Fat, Serum Lipoprotein Concentrations and Erythrocyte Membrane Fatty Acid Composition in Young Healthy Women (건강한 젊은 여성의 Apolipoprotein E 의 다형성 , 식이 지방 섭취, 혈청 지단백 농도와 적혈구 막의 지방산 조성과의 관계)

  • 박선민
    • Journal of Nutrition and Health
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    • v.30 no.8
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    • pp.936-951
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    • 1997
  • As fat consumption has increased in Korea , serum cholesterol concentrations have risen and the prevalence of atherosclerosis increased. The aim of the present study was to investigate the association among apolipoprotein E (Apo E) phenotypes, dietary fat consumption, plasma lipoprotein levels and fatty acid compositions of red blood cells in young healthy women. Seventy-one percent of participants had Apo E3/3, 14.9 percent had ApoE3/2 , 11.8% had Apo E4/3 and 1.2% had Apo E4.2. The subjects daily consumed approximately 1760kcal containing 29 energy % of fat. The ratio of polyunsaturated fat to saturated fat in a diet was 0. 73 . There were no significant differences of nutrient intakes according to Apo E phenotypes. Total cholesterol concentrations of subjects averaged approximately 160mg/dl , but 12 percent of them had over 200mg/dl, which is higher than the range recommended by the National Cholesterol Deucation Program. Notably, most subjects with 210mg/dl had Apo E4 isoforms. Subjects with Apo E4 isoforms had significantly higher total and LDL-cholesterol concentrations than those with Apo E3/2. Also subjects carrying Apo E4 isoforms tended to accumulated more fat in the body. Their BMI , WHR and arm fat area appeared to be how can arm fat area be greater no you mean grater, please check work vsage than subjects with Apo E3 isoforms, but not significantly. Fat intakes slightly influenced serum cholesterol levels. Myristic aicd intakes were positively correlated to serum total and LDL-choelsterol levels. Polyunsaturated fatty acid intakes were negatively correlated to serum total choelsterollevels. The results of 소냐 study suggest that , people with Apo E4 isoforms need to prevent the raising of serum total and LDL -cholesterol concentrations by reducing calorie and fat intakes and by maintaining a normal weight.

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Isolation, Molecular Phylogeny, and Tissue Distribution of Four cDNAs Encoding the Apolipoprotein Multigene Family in Barred Knifejaw, Oplegnathus fasciatus (Teleostei, Perciformes)

  • Kim, Keun-Yong;Cho, Young-Sun;Kim, Sung-Koo;Nam, Yoon-Kwon
    • Fisheries and Aquatic Sciences
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    • v.11 no.2
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    • pp.88-97
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    • 2008
  • Lipoproteins are complexes of lipids and specific apolipoproteins that are involved in lipid transport and redistribution among various tissues. In this study, we isolated full-length apolipoprotein cDNA sequences encoding apolipoprotein A-I (apoA-I), apoE, apoC-II, and apo-14 kDa in barred knifejaw, Oplegnathus fasciatus. In addition, we reconstructed phylogenetic trees and investigated mRNA tissue distributions. Alignment analyses of amino acid sequences revealed that secondary structures of the polypeptides apoA-I, apoE, and apoC-II in barred knifejaw are well conserved with their teleostean and mammalian counterparts in terms of characteristic tandem repetitive units forming amphipathic ${\alpha}$-helices. Both the sequence alignment data and cleavage sites of apo-14 kDa indicated a clear differentiation between Percomorpha and Cypriniformes. Meanwhile, the phylogenetic trees of apolipoprotein sub-families suggested that the common ancestor prior to the split of the Actinopterygii (ray-finned fishes) and Sarcopterygii (tetrapods) would have possessed the primordial protein-encoding genes. Tissue distribution of each apolipoprotein transcript determined by semi-quantitative RTPCR showed that barred knifejaw apoA-I transcripts were more or less ubiquitously expressed in the liver, intestines, brain, muscle, spleen, and kidney. The most striking difference from previous observations on barred knifejaw was the ubiquitous expression of apoE across all somatic tissues. Barred knifejaw apoC-II showed tissue-specific expression in the liver and intestines, while the liver and brain were the major sites of apo-14kDa mRNA synthesis.