• Preventive Nutrition and Food Science
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• 제4권2호
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• pp.139-144
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• 1999

Lipoprotein lipase (LPL) I san enzyme that catalyzed the hydrolysis of triacylglycerols of chylomicrons and VLDL to produce 20acylglycerols and fatty acids. The enzyme, LPL, is localized on the surface of the capillary endothelium and is widely distributed in extrahepatic tissues including heart, skeletal muscle and adipose tissue. LPL has been isolated from boving milk by affinity chromatography on heparin-separose in 2 M NaCL, 5mM barbital buffer, pH 7.4. To elucidate the lipid-binding regin, LPL was digested with trypsin and then separated by gel filtration. Lipid binding region of LPL has been investigated by recombining LPL peptides with DMPC vesicles. Proteolytic LPL fragments with DMPC were reassembled and stabilized by cholate. Lipid-binding region of LPL was identified by a PTH-automated protein sequencer, as AQQHYPVSAGYTK. The analysis of the secondary structure of the lipid-binding peptides revealed a higher probability of $\alpha$-helix structure compared to the whole LPL protein. The prediction of hydrophobicity of lipid -binding region was highly hydrophobic (-1.1) compared to LPL polypetide(-0.4).

• BMB Reports
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• 제42권1호
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• pp.1-5
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• 2009

Synaptic vesicles (SVs) are key structures for synaptic transmission in neurons. Numerous membrane-associated proteins are sorted from the Golgi complex to the axon and the presynaptic terminal. Protein-protein and protein-lipid interactions are involved with SV targeting in neurons. Interestingly, many SV proteins have lipid binding capability, primarily with either cholesterol or phosphoinositides (PIs). As examples, the major SV protein synaptophysin can bind to cholesterol, a major lipid component in SVs, while several other SV proteins, including synaptotagmin, can bind to PIs. Thus, lipid-protein binding plays a key role for the SV targeting of synaptic proteins. In addition, numerous SV proteins can be palmitoylated. Palmitoylation is thought to be another synaptic targeting signal. Here, we briefly describe the relationship between lipid binding and SV targeting.

• 분석과학
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• 제30권2호
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• pp.89-95
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• 2017

The thermodynamic functions for the binding of the peptide Substance P (SP) on the surface of lipid vesicles made of various types of lipids were obtained by using isothermal titration calorimetry. The reaction enthalpies measured from the experiments were -0.11 to $-4.5kcal\;mol^{-1}$. The sizes of the lipid vesicles were measured with dynamic light scattering instrument in order to get the correlation between the reaction enthalpies and the vesicle sizes. The bindings of SP on the lipid vesicles with diameter of 37 to 108 nm were classified into the enthalpy-driven reaction or the entropy-driven reaction according to the size of the lipid vesicles. For the enthalpy-driven binding reaction, the significance of the electrostatic interactions between SP and lipid molecules was affirmed from the experimental results of the DMPC/DMPG/DMPH and DMPC/DMPS/DMPH vesicles as well as the importance of the hydrophobic interactions between hydrophobic groups of SP and lipid molecules.

• 한국수산과학회지
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• 제37권5호
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• pp.359-365
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• 2004

To study the lipid adsorption characteristic of chitosans with different molecular weights and the degrees of deacetylation, in vitro test and near-infrared (NIR) spectroscopic analysis have been performed for the measurement of lipid adsorption characteristics of chitosan. The degrees of deacetylation in chitosans were $70{\%},\;85{\%}\;and\;92{\%}$ at different deacetylation times (1 hr, 2 hrs, 3 hrs), respectively. The molecular weight of each chitosan was controlled by enzymatic hydrolysis, and then the molecular weight of the chitosan was 4 kDa. The bulk density, water holding capacity and fat binding capacity of each chitosan powder were $96.2-504.0{\%},\;374.4-1217.9{\%},\;and\;307.0-659.3{\%}$, respectively. The higher molecular weight of chitosan was exhibited the lower bulk density and the higher water and fat binding capacities. Bindinf capacities of chitosan powders to bile salts, cholesterol and linoleic acid were $41.2-63.3{\%},\;40.8-67.4{\%},\;42.6-72.6{\%}$, respectively. In NIR spectrum of lipid adsorbed chitosan the occurrence static eletronical binding between chitosan and lipid was identified by NIR spectrum peak induced from combination of carboxylic group in lipid and amino group in chitosan. In conclusion, the higher degree of deacetylation and molecular weight of chitosan showed the higher lipid binding capacity and the lipid adsorption of chitosan were occurred by combination of carboxylic group in lipids and amino group in chitosan.

• Journal of Nutrition and Health
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• 제32권6호
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• pp.628-636
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• 1999

There is a marked increase in geriatric disease, especially liver disease, due to the continuous increase in alcohol and fat consumption. Since the fatty liver, induced by alcohol or fat, is basically from abnormalities in the lipid metabolism, it is possible that fatty acid binding protein(FABP) which is related to the fatty acid metabolism may also be abnormal in these livers. FABP is a small molecular weight protein family present in cytosol in high concentration. It has been proposed as a fatty acid transfer protein and as a binding protein responsible for controlling intracellular free fatty acid concentration. In this research, we have examined the relationship between liver FABP and fatty liver induced by alcohol or high cholesterol diet. Rats were fed one of either semipurified liquid diets; control diet containing 65% carbohydrate, 20% protein, and 15% fat or high cholesterol diet containing 1%(w/w) cholesterol or alcohol diet containing 37% of alcohol instead of carbohydrate. After 5 weeks of feeding period, all rats received commercial chow diet for 5 weeks to examine recovery effect. Liver and blood samples were collected at 0, 1, 3, 5 and 10 weeks to analyze lipid compositions. FABP was purified from liver cytosol and injected to rabbit to obtain antiserum. Liver FABP amount was determined by SDS-PAGE and western blotting methods. Fatty acid binding capacity was determined by binding of 14Cpalmitate with the delipidated liver cytosol. Consumption of alcohol increased serum cholesterol, triglyceride concentration and decreased HDL-cholesterol concentration after 5 weeks. Serum apolipoprotein B concentration increased after 3 weeks and LDL-cholesterol and apolipoprotein A concentration changed after 1 week. Liver cholesterol and triglyceride concentration increased after 3 weeks. Consumption of high cholesterol diet changed liver and serum lipid composition after 3 weeks. Swiching to normal diet for 5 weeks did not normalize most of lipid composition in serum and liver except serum and liver except serum cholesterol, triglyceride and liver cholesterol. Liver cytosol FABP content and the fatty acid binding capacity decreased dramatically after 1 week with alcohol consumption. This results indicate that FABP content changes before the changes before the changes of blood or liver lipid composition, suggesting changes of FABP may cause development of the fatty liver induced by alcohol and can be used as an index of detecting a early development of fatty liver.

• BMB Reports
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• 제42권6호
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• pp.367-372
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• 2009

In this study, we showed that WAX9D, a nonspecific lipid-transfer protein found in broccoli, binds palmitate (C16) and stearate (C18) with dissociation constants of 0.56 ${\mu}M$ and 0.52 ${\mu}M$, respectively. WAX9D was fused to thioredoxin protein by genetic manipulation to enhance its solubility. The data revealed strong interaction of Trx-WAX9D with palmitate and stearate. The dissociation constants of Trx-WAX9D for palmitate and stearate were 1.1 ${\mu}M$ and 6.4 ${\mu}M$, respectively. The calculated number of binding sites for palmitate and stearate was 2.5 to 2.7, indicating that Trx-WAX9D can bind three molecules of fatty acids. Additionally, Trx-WAX9D was shown to inhibit the apoptotic effect of palmitate in endothelial cells. Our data using Trx-WAX9D provide insight into the broad spectrum of its biological applications with specific palmitate binding.

• The Korean Journal of Physiology and Pharmacology
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• 제23권3호
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• pp.161-169
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• 2019

Fumigaclavine C (FC), an active indole alkaloid, is obtained from endophytic Aspergillus terreus (strain No. FC118) by the root of Rhizophora stylosa (Rhizophoraceae). This study is designed to evaluate whether FC has anti-adipogenic effects in 3T3-L1 adipocytes and whether it ameliorates lipid accumulation in high-fat diet (HFD)-induced obese mice. FC notably increased the levels of glycerol in the culture supernatants and markedly reduced lipid accumulation in 3T3-L1 adipocytes. FC differentially inhibited the expressions of adipogenesis-related genes, including the peroxisome proliferator-activated receptor proteins, CCAAT/enhancer-binding proteins, and sterol regulatory element-binding proteins. FC markedly reduced the expressions of lipid synthesis-related genes, such as the fatty acid binding protein, lipoprotein lipase, and fatty acid synthase. Furthermore, FC significantly increased the expressions of lipolysis-related genes, such as the hormone-sensitive lipase, Aquaporin-7, and adipose triglyceride lipase. In HFD-induced obese mice, intraperitoneal injections of FC decreased both the body weight and visceral adipose tissue weight. FC administration significantly reduced lipid accumulation. Moreover, FC could dose-dependently and differentially regulate the expressions of lipid metabolism-related transcription factors. All these data indicated that FC exhibited anti-obesity effects through modulating adipogenesis and lipolysis.

• 대한약리학회지
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• 제21권2호
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• pp.79-89
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• 1985

심근 세포의 칼슘 조절에 중요한 역할을 하는 sarcoplasmic reticulum (SR)의 칼슘운반 능력이 허혈 심근에서 현저히 억제됨이 알려져 있다. 이와같은 허혈 심근에서의 SR 칼슘운반승력 저하에 유독성 산소 대사물이 관여할 것으로 생각되고 있으나 그 기전에 관하여는 아직 알려진 바 없다. 본 연구에서는 그 기전의 일단을 규명하기 위하여 산틴 산화효소계에 의하여 발생된 유독성 산소대사물긴 돼지 심실근에서 추출한 sarcoplasmic reticulum의 칼슘흡수 및 막지질 과산화, sulfhydryl group 그리고 단백질 변성에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1) 산틴 산화 효소계와 반응시킨 sarcopl smic reticulum의 칼슘흡수는 반응시간 경과에 따라 현저히 억제되었다. 2) sarcoplasmic reticulum 막지질 과산화는 산딘 산화 효소계에 의하여 현저히 증가되었다 3) 항산화제 ${\beta}$-phenylenediamine은 막지질 과산화의 증가는 효과적으로 억제하였으나, 칼슘흡수 억제는 부분적으로 회복시켰다. 4) 산틴 산화 효소계에 의하여 SH-group은 현저히 감소되었으며, 항산화제 첨가에 의하여 그 감소가 일부 억제되었다. 5) sarcoplasmic reticulum을 DTNB로 처리하여 SH-group을 산소 대사물에 의한 산화반응으로부터 보호했을 경우 칼슘흡수의 억제가 부분적으로 방지되었다. 6) Sephadex G-200 크로마토그라피 상에서 산틴 산화효소계와 반응시킨 sarcoplasmic reticulum의 단백질분해가 관찰되었다. 7) 단백질의 polymerization은 관찰되지 않았으며, 아울러 polymerization을 억제하는 semicarbazide로 칼슘흡수 감소를 방지하지 못하였다. 이상의 결과에서 유독성 산소대사물에 의한 sarcoplasmic reticulum의 칼슘흡수 억제는 sarcoplasmic reticulum의 막지질 과산화, SH-group의 산화 및 막 반백절의 분해 등으로 초래되는 복합적인 기전으로 추정되었다.

• BMB Reports
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• 제57권2호
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• pp.92-97
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• 2024

Elevated blood glucose is associated with an increased risk of atherosclerosis. Data from the current study showed that glucosamine (GlcN), a normal glucose metabolite of the hexosamine biosynthetic pathway (HBP), promoted lipid accumulation in RAW264.7 macrophage cells. Oleic acid- and lipopolysaccharide (LPS)-induced lipid accumulation was further enhanced by GlcN in RAW264.7 cells, although there was no a significant change in the rate of fatty acid uptake. GlcN increased acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), scavenger receptor class A, liver X receptor, and sterol regulatory element-binding protein-1c (SREBP-1c) mRNA expression, and; conversely, suppressed ATP-binding cassette transporter A1 (ABCA-1) and ABCG-1 expression. Additionally, GlcN promoted O-GlcNAcylation of nuclear SREBP-1 but did not affect its DNA binding activity. GlcN stimulated phosphorylation of mammalian target of rapamycin (mTOR) and S6 kinase. Rapamycin, a mTOR-specific inhibitor, suppressed GlcN-induced lipid accumulation in RAW264.7 cells. The GlcN-mediated increase in ACC and FAS mRNA was suppressed, while the decrease in ABCA-1 and ABCG-1 by GlcN was not significantly altered by rapamycin. Together, our results highlight the importance of the mTOR signaling pathway in GlcN-induced macrophage lipid accumulation and further support a potential link between mTOR and HBP signaling in lipogenesis.

• Bulletin of the Korean Chemical Society
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• 제34권3호
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• pp.743-748
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• 2013

The temperature dependence of the partition coefficients of a neuropeptide, substance P (SP), in isotropic acidic bicelles was investigated by using a pulsed field gradient nuclear magnetic resonance diffusion technique. The addition of negatively charged dimyristoylphosphatidylserine to the neutral bicelle changed the SP partitioning a little, which implies that the hydrophobic interaction between the hydrophobic residues of SP and the acyl chains of lipid molecules is the major interaction while the electrostatic interaction is minor in SP binding in a lipid membrane. From the temperature dependence of the partition coefficients, thermodynamic functions were calculated. The partitioning of SP into the acidic bicelles is enthalpy-driven, as it is for small unilamellar vesicles and dodecylphosphocholine micelles, while peptide partitioning into a large unilamellar vesicle is entropy-driven. This may mean that the size of lipid membranes is a more important factor for peptide binding than the surface curvature and surface charge density.