• Journal of Microbiology and Biotechnology
• /
• 제24권12호
• /
• pp.1744-1754
• /
• 2014

The hepatitis C virus (HCV) RNA genome is replicated by an RNA replicase complex (RC) consisting of cellular proteins and viral nonstructural (NS) proteins, including NS5B, an RNA-dependent RNA polymerase (RdRp) and key enzyme for viral RNA genome replication. The HCV RC is known to be associated with an intracellular membrane structure, but the cellular components of the RC and their roles in the formation of the HCV RC have not been well characterized. In this study, we took a proteomic approach to identify stomatin, a member of the integral proteins of lipid rafts, as a cellular protein interacting with HCV NS5B. Co-immunoprecipitation and co-localization studies confirmed the interaction between stomatin and NS5B. We demonstrated that the subcellular fraction containing viral NS proteins and stomatin displays RdRp activity. Membrane flotation assays with the HCV genome replication-competent subcellular fraction revealed that the HCV RdRp and stomatin are associated with the lipid raft-like domain of membranous structures. Stomatin silencing by RNA interference led to the release of NS5B from the detergent-resistant membrane, thereby inhibiting HCV replication in both HCV subgenomic replicon-harboring cells and HCV-infected cells. Our results identify stomatin as a cellular protein that plays a role in the formation of an enzymatically active HCV RC on a detergent-resistant membrane structure.

• 생명과학회지
• /
• 제11권2호
• /
• pp.103-110
• /
• 2001

This study was designed to investigate the effects of silkworm(Bombyx moril L.) powder on oxidative stress and membrane fluidity in brain membranes of rats. Sprague-Dawley(SD) male rats(160$\pm$10 g) were fed basic diet(control group), and experimental diets(SWP-200 and SWP-400 groups) added 200 and 400mg/kg BW/day for 6 weeks. There were no significant differences in cholesterol levels of brain memberanes by administration of silkworm powder (SWP). Membrane fluidities were significantly increased(21.5% and 30.8%, respectively) in brain mitochondria of SWP-200 and SWP-400 groups compared with control group, but significant difference between brain microsomes could not obtained. Basal oxygen radicals (BORs) in brain mitochondria and mircrosomes were significantly inhibited(8.5% and 16.5%, 16.8%and 24.8%, respectively) by SWP-200 and SEP-400 groups compared with control group. Induced oxygen radicals(IORs) in brain mitochondria were significantly inhibited(16.6% and 21.4%, respectively)by sWP-200 and SWP-400 groups compared with control group, but IOR in brain microsome were significantly inhibited about 16.0% by SWP-400 groups only compared with control group. Lipid peroxide(LPO) levels were significantly decreaed(14.8%and 22.4%, respectively) in brain mitochondria of SWP-200 and SWP-400 groups compared with control group, but LPO level was significantly decreased about 16.0% in brain microsome of SWP-400 group only. Oxidized protein(OP) levels were remarkably decreased(about 14.8% and 16.5%, respectively) in brain mitochondria of SWP-200 and SWP-400 groups, but OP level was significantly decreased about 13.0% in brain microsome of SWP-400 group only compared with control group, Theses results suggest that administration of in brain microsome of SWP-400 group only compared with control group. These results suggest that administration of SWP may play effective role in attenuating an oxidative stress and increasing a membrane fluidity in brain membranes.

• 생명과학회지
• /
• 제10권4호
• /
• pp.354-361
• /
• 2000

The effect of mulberry (Morus alba L.) leaf extract(MLE) on oxidative stress and membrane fluidity in brain membranes of SD rats fed with 100 and 300 mg/kg BW/day were carried out for 6 weeks. Cholesterol accumulations resulted in a consistent decreases (4.6% and 5.6%, respectively) in brain mitochondria and microsomes of MLE-300 group compared with control group. Membrane fluidities were dose-dependently increased (2.2% and 5.1%, 5.0% and 15.2%) in brain mitochondria and microsomes of MLE-100 and MLE-300 groups compared with control group. Basal oxygen radicals(BORs) in brain mitochondria and microsomes were significantly inhibited (15.7% and 25.1%, 9.0% and 12.4%, respectively) by MLE-100 and MLE-300 groups compared with control group. Induced oxygen radicals(IORs) in brain mitochondria and microsomes were significantly inhibited (8.9% and 13.1%, 16.5% and 23.2%, respectively) by MLE-100 and MLE-300 groups compared with control group. Lipid peroxide (LPO) levels were significantly decreased (8.5% and 18.1%, 7.6% and 12.3%) in brain mitochondria and microsomes of MLE-100 and MLE-300 groups compared with control group. Oxidized protein (OP) levels were dose-dependently decreased (4.3% and 14.2%, 10.0% and 10.9%, respectively) in brain microsomes of MLE-100 and MLE-300 groups compared with control group. These results suggest that MLE may play an effective role in an attenuating an oxidative stress and increasing a membrane fluidity in brain membranes.

• Preventive Nutrition and Food Science
• /
• 제3권3호
• /
• pp.211-215
• /
• 1998

Preparative isolation of lipid granules from Fhodotorula glutinis, which has been studied for long time to produce edible lipids, was carried out by flotation method in Ficoll-Linear density gradient. When the isolated lipid granules were suspended in a series of solutions containing varying concentration of osmotic stabilizer (sorbitoal and mannitol) ranging from 0.8M to 0M, the lipid granules appeared to be disrupted at a concentration between 0.8M and 0.7, and again at a concentration below 0.1M, suggesting that lipid granules have a membraneous structure and that at least two types of lipid granules are present. Compositional analysis of lipids from lipid granules revealed that lipids are composed mainly of neutral lipids (87.8% of total lipids), predominantly as triacylglycerols (71.89%). Marked differences were observed inphospholipids between lipids of lipid granules and those of whole cells . The major components of phospholipids in lipid granules and inwhole cells are phosphatidylcholine(38.6%) and phosphatidylserine(42.8%), respectively. In addition, significant differences were also observed in the fatty acid composition of phospholipids. As phospholipids are important structural components of membranes, these differences lead to the suggesting that the membrane of lipid granules may be distinct functionally and structurally from other membranes of yeast cells. The major fatty acid components of neutral lipidss of whole cells and lipid granules are palmitic , oleic and linoleic acid. However , degreeof fatty acid unsaturation of neutal lipids of lipid granules was much lower than that of neutral lipids of whole cells.

• Bulletin of the Korean Chemical Society
• /
• 제33권2호
• /
• pp.426-432
• /
• 2012

The activity of an antimicrobial peptide, protegrin-1 (PG-1), on lipid membranes was investigated using solidstate NMR and a new sampling method that employed mechanically aligned bilayers between thin glass plates. At 95% hydration and full hydration, the peptide respectively disrupted 25% and 86% of the aligned 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphotidylcholine (POPC) bilayers at a P/L (peptide-to-lipid) ratio of 1/20 under the new experimental conditions. The kinetics of the POPC bilayers disruption appeared to be diffusioncontrolled. The presence of cholesterol at 95% hydration and full hydration reduced the peptide disruption of the aligned POPC bilayers to less than 10% and 35%, respectively. A comparison of the equilibrium states of heterogeneously and homogeneously mixed peptides and lipids demonstrated the importance of peptide binding to the biomembrane for whole membrane disruption.

• BMB Reports
• /
• 제32권5호
• /
• pp.440-444
• /
• 1999

Membrane lipid peroxidation processes yield reactive aldehydes that may react with copper,zinc superoxide dismutase (Cu,Zn SOD), one of the key antioxidant enzymes against oxidative stress. We investigated this possibility and found that exposing Cu,Zn SOD to malondialdehyde (MDA) or 4-hydroxynonenal (HNE) caused the loss of dismutase activity, cross-linking of peptides, and an increase in protein oxidation, reflected by the increased level of carbonyl groups. When Cu,Zn SOD that had been exposed to MDA or HNE was subsequently analyzed by amino acid analysis, histidine content was found to be significantly lost. Both MDA-and HNE-treated Cu,Zn SOD were resistant to proteolysis, which may imply that damaged proteins exist in vivo for a longer period of time than the native enzyme. The lipid peroxidation-mediated damage to Cu,Zn SOD may result in the perturbation of cellular antioxidant defense mechanisms, and subsequently lead to a pro-oxidant condition.

• 약학회지
• /
• 제38권6호
• /
• pp.786-790
• /
• 1994

The crude alkaloidal fraction of the root of Cynanchum caudatum Max.(Asclepiadaceae) was tested for the effects on the activities of free radical generating enzymes and the formation of lipid peroxide. Aldehyde oxidase was strongly inhibited to about 90% of the activity by treating 1.0 mg/ml of alkaloidal fraction, corresponding to competitive inhibition. Moreover, the formation of lipid peroxide which causes damage of cell membrane was reduced in proportion to the increasing alkaloid concentration. However, xanthine oxidase of which structure and function are similar to those of aldehyde oxidase was not inhibited by the alkaloidal fraction.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
• /
• pp.381.2-381.2
• /
• 2002

There is now increasing evidence that free radicals and active oxygen species are involved in a variety of pathological events. often associated with ageing. Free radical-mediated cell damage and free radical attack on polyunsaturated fatty acids result in the formation of lipid radicals. These lipid radicals react readily with molecular oxygen to produce peroxy radicals responsible for initiating lipid peroxidation. The peroxidation of cellular membrane lipid can lead to cell necrosis and considered to be implicated in a number of pathophysiological conditions as well as in the toxicity of many xenobiotics. (omitted)

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
• /
• pp.263.2-263.2
• /
• 2003

There is now increasing evidence that free radicals and active oxygen species are involved in a variety of pathological events. Free radical-mediated cell damage and free radical attack on polyunsaturated fatty acids result in the formation of lipid radicals. These lipid radicals react readily with molecular oxygen to produce peroxy radicals responsible for initiating lipid peroxidation. The peroxidation of cellular membrane lipid can lead to cell necrosis and considered to ve implicated in a number of pathophysiological conditions including liver disease. (omitted)

• Journal of Microbiology and Biotechnology
• /
• 제32권1호
• /
• pp.37-45
• /
• 2022

The fungal cell wall and membrane are the principal targets of antifungals. Herein, we report that myricetin exerts antifungal activity against Candida albicans by damaging the cell wall integrity and notably enhancing the membrane permeability. In the presence of sorbitol, an osmotic protectant, the minimum inhibitory concentration (MIC) of myricetin against C. albicans increased from 20 to 40 and 80 ㎍/ml in 24 and 72 h, respectively, demonstrating that myricetin disturbs the cell wall integrity of C. albicans. Fluorescence microscopic images showed the presence of propidium iodide-stained C. albicans cells, indicating the myricetin-induced initial damage of the cell membrane. The effects of myricetin on the membrane permeability of C. albicans cells were assessed using crystal violet-uptake and intracellular material-leakage assays. The percentage uptakes of crystal violet for myricetin-treated C. albicans cells at 1×, 2×, and 4× the MIC of myricetin were 36.5, 60.6, and 79.4%, respectively, while those for DMSO-treated C. albicans cells were 28.2, 28.9, and 29.7%, respectively. Additionally, myricetin-treated C. albicans cells showed notable DNA and protein leakage, compared with the DMSO-treated controls. Furthermore, treatment of C. albicans cells with 1× the MIC of myricetin showed a 17.2 and 28.0% reduction in the binding of the lipophilic probes diphenylhexatriene and Nile red, respectively, indicating that myricetin alters the lipid components or order in the C. albicans cell membrane, leading to increased membrane permeability. Therefore, these data will provide insights into the pharmacological worth of myricetin as a prospective antifungal for treating C. albicans infections.