• Archives of Pharmacal Research
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• v.9 no.2
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• pp.75-79
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• 1986

The effect of phenothiazine derivatives on the thermotropic transition of liposomal lipid bilayer made of dipalmitoyl phosphatidylchline and dipalmitoyl phosphatidic acid was investigated with differential scanning calorimetry. The thermograms of the liposomal bilayer incorporated with levomepromazine, chlopromazine, prochloperazine, perphenazine and fluphenazine were obtained and the size of cooperative unit of the transition were calculated from the ratio of the van't Hoff enthalpy change to the calculated enthalpy change of the transition. The results showed that incorporation of phenothiazine derivatives into the liposomal bilayer reduced the transition temperature at which the transition from solid state to liquid-crystalline state occurs, and broadened the thermogram peaks. Phenothiazine derivatives also significantly reduced the size of cooperative unit of the transition. The effect of the drugs was proportional to the concentration of the drug in the bilayer. This means that phenothiazine derivatives might have significant fluidizing effects on the biomembrane. The sizes of cooperative unit were successfully corrlated with phar-macological activities of the drugs and the surface pressure increases of lipid monolayer by these drugs. These correlations might be ascribed to a possible hydrophobic nature of interaction between the biomembrane and the drugs involved in their pharmacology.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.3
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• pp.323-327
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• 1993

The effect of soyasaponin on the liposomal phospholipid membrane was investigated by differential scanning calorimetry (DSC). Soyasaponins were obtained and the enthalpy changes and the sizes of cooperative unit of the transition were calculated. The thermograms of L-$\alpha$-dimyristoyl phosphatidylcholine (DMPC) incorporated with soyasaponin showed that the phase transition temperature was significantly lowered and the peak was broadened. This was attributed to the possibility that incorporation of soyasaponin into the lipid bilayers reduced the cooperative unit of phospholipid bilayers. These results indicate soyasaponin might have significant effect on the fluidity of biological membrane.

• KSBB Journal
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• v.7 no.4
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• pp.302-307
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• 1992

Protein phosphatase 2A was obtained from a cytosolic fraction of bovine brain homogenate. The phosphatase activity using phosphorylated histone Hl as substrate was suppressed in the presence of liposomes composed of dipalmitoylphosphatidylcholine(DPPC) or the mixture of phosphatidylserine and DPPC. The binding of protein phosphatase to liposome was indicated by the facts that the phosphatase activity of the supernatant of protein phosphatase/multilayer vesicle mixture was decreased with increasing amount of liposome, and that [$^{125}I$]-labeled protein phosphatase was coeluted with liposome. However, the affinity of the protein for phospholipid membrane was not so high. On the other hand, okadaic acid and liposome reduced the phosphatase activity synergistically, which means that okadaic acid binds neither to lipid membrane nor to the membrane-associated phosphatase, The inhibitory effect of liposome was, therefore, ascribed to association of the protein phosphatase 2A with the lipid bilayer membrane.

• Bulletin of the Korean Chemical Society
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• v.11 no.2
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• pp.131-134
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• 1990

Cytochrome c induces fusion of phosphatidylserine /phosphatidylethanolamine vesicles while apocytochrome c does not have a fusogenic capability despite the fact that the apoprotein binds to the vesicles more extensively. In order to see whether the difference in the fusogenic behavior comes from the topological variation in membrane bound proteins, the holoprotein and apoprotein were labeled with phenylisothiocyanate, a hydrophobic label, in the presence of its hydrophilic analogue p-sulfophenylisothiocyanate. Apocytochrome c was labeled with the hydrophobic probe more extensively than the cytochrome c, indicating that the apoprotein penetrates deeper into the bilayer than cytochrome c does. The translocation experiments of these proteins by trypsin entrapped vesicles further supported this conclusion.

• Bulletin of the Korean Chemical Society
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• v.17 no.3
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• pp.239-244
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• 1996

Mastoparan B (MP-B) that is a novel MP isolated from the hornet Vespa basalis, was studied as compared with MP, in terms of interaction with phospholipid bilayer and antimicrobial activity. MP-B has more hydrophilic amino acid residues in hydrophilic face of amphiphilic α-helical structure than MP. The both peptides exhibited considerably different effect on interaction with lipid bilayers, e.g. their conformation in the presence of acidic and neutral liposomes, dye-release ability from encapsulated liposomes, but on the whole the interaction mode was similar. On antimicrobial activity, MP had a strong activity against Gram-positive bacteria but no against Gram negative ones. Contrary to this, MP-B had a strong activity against Gram-positive and potent against Gram-negative ones. Since both peptides have almost same residues on the hydrophobic side, such more hydrophilic surface on the molecule seems to lead to the subtle change in its interaction with membranes, resulting in the alternation in its biological activity.

• BMB Reports
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• v.34 no.5
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• pp.446-451
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• 2001

The interaction of cytochrome c with binary phospholipid mixtures was investigated by solid-state $^2H$- and $^{31}P$-NMR. To examine the effect of the interaction on the glycerol backbones, the glycerol moieties of phosphatidylcholine (PC), and cardioliph (CL) were specifically deuterated. On the binding of cytochrome c to the binary mixed bilayers, no changes in the quadrupole splittings of each of the components were observed for the PC/PG, PE/CL and PE/PG liposomes. In contrast, the splittings of CL decreased on binging of protein to the PC/CL liposomes, although those of PC did not change at all. This showed that cytochrome c specifically interacts with CL in PC/CL bilayers, and penetrates into the lipid bilayer to some extent so as to perturb the dynamic structure of the glycerol backbone. This is distinctly different from the mode of interaction of cytochrome c with other binary mixed bilayers. In the $^{31}P$-NMR spectra, line broadening and a decrease of the chemical shift anisotropy were observed on the binding of cytochrome c for all binary mixed bilayers that were examined. These changes were more significant for the PC/CL bilayers. Furthermore, the line broadening is more significant for PC than for CL in PC/CL bilayers. Therefore, it can be concluded that with the polar head groups, not only CL but also PC are involved in the interaction with cytochrome c.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.4
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• pp.459-468
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• 1984

The currently accepted model of membrane structure proposes a dynamic, asymmetric lipid matrix of phospholipids and cholesterol with globular proteins embedded across the membrane to various degrees. Most phospholipids are in the bilayer arrangement and also closely associated with integral membrane proteins or loosely associated with peripheral proteins. Biological functions of membrane, such as membrane-bound enzyme functions and transport systems, are influenced by the membrane physical properties, which are determined by fatty acid composition of phospholipids, polar head group composition and membrane cholesterol content. Polar and non-polar region of the phospholipid molecule can interact, with changes in the conformation of a membrane-associated protein altering either its catalytic activity or the protein's interaction with other membrane proteins. Mammalian dietary studies attempted to change the lipid composition of a few cell membranes have shown comparisons, using essential fatty acid-deficient diets. In recent years, Clandinin and a few other workers have pioneered the study proving the influence of dietary fat fed in a nutritionally complete diet on composition of phospholipid classes of cell membrane. Modulation caused by diet fat was rapid and reversible in phospholipid fatty acyl composition of membranes of cardiac mitochondria, liver cell, brain synaptosome and lymphocytes. These changes were at the same time, accompanied by variety of membrane associated functions controlled by membrane-bound enzymes, tranporter and receptor proteins. The findings suggest the basic concept of the necessity of dietary fatty acid balance if consistency of optimal membrane structural lipid composition is to be maintained, as well as the overall inadequacy of describing the nutritional-biochemical quality of a dietary fat solely by its content of linoleic acid. Furthermore, they give light on the possible application to clinical and preventive medicine.

• KSBB Journal
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• v.6 no.3
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• pp.281-288
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• 1991

This research was carried to investigate the effects of several organic solvents on the enzymatic transphosphatidylation in emulsion and two-phase solvent systems. The solvents having a similar dielectric constant with diethylether were effective for the enzyme activity. Diethylether and butylacetate were the most effective solvents, when added 12-15%(v/v) and 10-40%(v/v), respectively, for the synthesis of phosphatidylglycerol, phosphatidylethyleneglycol and phosphatidylpropyleneglycol. In the emulsion system, the size of ovolecithin liposome was increased and the clearness of the phospholipid bilayer was reduced as increasing the diethylether concentration. In the twophase solvent system, the rapidest reaction was obtained when water-organic solvent ratio was close to 1. The ratio of aqueous phase. however, should be lowered to 37% to gain the sole product of transphosphatidy1ation, without phosphatidohydrolysis.

• BMB Reports
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• v.32 no.6
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• pp.561-566
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• 1999

CA(1-8)-ME(1-12) [CA-ME], composed of cecropin A(1-8) and melittin(1-12), is a synthetic antimicrobial peptide having potent antibacterial and antitumor activities with minimal hemolytic activity. In order to investigate the effects of the flexible hinge sequence, Gly-Ile-Gly, of CA-ME on antibiotic activity, CA-ME and three analogues, CA-ME1, CA-ME2, and CA-ME3, were synthesized. The Gly-Ile-Gly sequence of Ca-ME was deleted in CA-ME1 and replaced with Pro and Gly-Pro-Gly in CA-ME2 and CA-ME3, respectively. CA-ME1 and CA-ME3 showed a significant decrease in antitumor activity and phospholipid vesicle-disrupting ability. However, CA-ME2 showed similar antitumor and vesicle-disrupting activities, as compared with CA-ME. These results suggest that the flexibility or ${\beta}$-turn induced by Gly-Ile-Gly or Pro in the central part of CA-ME may be important in the electrostatic interaction of the N-terminus cationic ${\alpha}$-helical region with the cell membrane surface and the hydrophobic interaction of the C-terminus amphipathic ${\alpha}$-helical region with the hydrophobic acyl chains in the cell membrane. CA-ME3 exhibited lower antitumor and vesicle-disrupting activities than CA-ME and CA-ME2. This result suggests that the excessive ${\beta}$-turn structure caused by the Gly-Pro-Gly sequence in CA-ME3 seems to interrupt ion channel/pore formation in the lipid bilayer. We concluded that the appropriate flexibility or bilayer. We concluded that the appropriate flexibility or ${\beta}$-turn structure provided by the central hinge is responsible for the effective antibiotic activity of the antimicrobial peptides with the helix-hinge-helix structure.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.1
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• pp.39-47
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• 2009

Gaegurin 4(GGN 4), an antimicrobial peptide isolated from a Korean frog, is five times more potent against Gram-positive than Gram-negative bacteria, but has little hemolytic activity. To understand the mechanism of such cell selectivity, we examined GGN4-induced $K^+$ efflux from target cells, and membrane conductances in planar lipid bilayers. The $K^+$ efflux from Gram-positive M. luteus(2.5 ${\mu}g/ml$) was faster and larger than that from Gram-negative E. coli(75 ${\mu}g/ml$), while that from RBC was negligible even at higher concentration(100 ${\mu}g/ml$). GGN4 induced larger conductances in the planar bilayers which were formed with lipids extracted from Gram-positive B. subtilis than in those from E. coli(p<0.01), however, the effects of GGN4 were not selective in the bilayers formed with lipids from E. coli and red blood cells. Addition of an acidic phospholipid, phosphatidylserine to planar bilayers increased the GGN4-induced membrane conductance(p<0.05), but addition of phosphatidylcholine or cholesterol reduced it(p<0.05). Transmission electron microscopy revealed that GGN4 induced pore-like damages in M. luteus and dis-layering damages on the outer wall of E. coli. Taken together, the present results indicate that the selectivity of GGN4 toward Gram-positive over Gram-negative bacteria is due to negative surface charges, and interaction of GGN4 with outer walls. The selectivity toward bacteria over RBC is due to the presence of phosphatidylcholine and cholesterol, and the trans-bilayer lipid asymmetry in RBC. The results suggest that design of selective antimicrobial peptides should be based on the composition and topology of membrane lipids in the target cells.