• Bulletin of the Korean Chemical Society
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• v.27 no.3
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• pp.386-388
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• 2006

Membrane proteins in highly oriented lipid bilayer samples are useful for membrane protein structure determination. We used in the past planar lipid bilayers which were aligned and supported on the glass slide. These samples were mechanically aligned in a magnetic field. However, these stacks of glass slides with planar lipid bilayers are not well suited for use with a commercial solid-state NMR probe with a round coil. Therefore, a homebuilt solid-state NMR probe was built and used with a stack of thin glass plates wherein the RF coil was wrapped directly around the flat square sample. Recently, we began to use magnetically aligned bicelles that are suitable for the structure determination of membrane proteins by solid-state NMR spectroscopy without any effort to build a flat square coil probe. These bicelle samples are well suited for use with a commercial solidstate NMR probe with a round coil, are very easy to prepare and are very stable, so that they can be kept for more than a year. In this paper, we present the solid-state NMR spectra of optimized and magnetically oriented bicelle samples of membrane proteins.

• Proceedings of the Korean Biophysical Society Conference
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• 1998.06a
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• pp.16-16
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• 1998

The role of phospholipids in the membrane binding and subsequent insertion of the microsomal protein rabbit cytochrome P450 (P450) lA2 into phospholipid bilayers was investigated. The insertion of P450 lA2 into phospholipid bilayers was determined by the amount of quenching of Trp fluorescence of P450 lA2 by pyrene and brominated and doxyl-labeled phospholipids.(omitted)

• Bulletin of the Korean Chemical Society
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• v.18 no.9
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• pp.933-938
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• 1997

The interaction of mastoparan B, a tetradecapeptide toxin found in the hornet Vespa basalis, with phospholipid bilayers was investigated. Synthetic mastoparan B and its analogs, obtained by substituting one hydrophilic amino acid (2-Lys, 4-Lys, 5-Ser, 8-Ser, 11-Lys, or 12-Lys) in mastoparan B with Ala, were studied. Mastoparan B and its analogs were synthesized by the solid-phase method. As shown by circular dichroism spectra, mastoparan B and its analogs adopted an unordered structure in buffer solution. All peptides took an α-helical structure, and the α-helical content of its analogs increased in the presence of neutral and acidic liposomes as compared to that of mastoparan B. In the calcein leakage experiment, we observed that mastoparan B interacted more weakly with lipid bilayers in neutral and acidic media than its analogs. Mastoparan B also showed slightly lower antimicrobial activity and hemolytic activity towards human erythrocytes than its analogs. These results indicate that the greater hydrophobicity of the amphiphilic α-helix of mastoparan B by replacement with alamine residues results in the increased biological activity and helical content.

• 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.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.

• Journal of fish pathology
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• v.9 no.1
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• pp.65-77
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• 1996

Tachyplesin I is an antimicrobial peptide isolated from horseshoe crab. To investigate the mechanism of action of tachyplesin I for phospholipid bilayers, tachyplesin I and five analogs have been synthesized by the solution method. The synthesized five analogs are [$Phe^2$]-tachyplesin I, [$Phe^{8,13}$]-tachyplesin I, [$Cys(Acm)^{3,7,12,16}$]-tachyplesin I with no disulfide bonds, 7(Acm) and 10 (Acm) which denote the fragments [$Cys(Acm)^{3,7,12,16}$]-tachyplesin I. Circular dichroism spectra showed that tachyplesin I took an antiparallel $\beta$-structure in buffer solution and a less ordered structure in acidic liposomes. The carboxyfluorescein leakage experiment indicated that tachyplesin I interacted strongly with neutral and acidic phospholipid bilayers. In fluorescence experiment, the hydrophobic part of the peptide was shown to be embedded in lipid bilayers. All the peptides except for 7(Acm) and 10(Acm) were almost equally active in lipopolysaccharide binding. Therefore, the present study suggested that phospholipid bilayers induced a conformational change of tachyplesin I from the stable $\beta$-structure to a less ordered one.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.3
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• pp.518-524
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• 1998

Liposomes have been widely employed as biomembrane-mimetic system and drug-delivery system. In these applications, the low stability of liposomes has been the most serious problem. They have relatively short half-lives and easily lysed through interactions with biological components. This study was performed to investigate the effects of godulbaegi extracts on the fludity of phospholipid liposomes. We used dipalmitoyl phosphatidylcholine(DPPC) liposomes which make most stable liposomes among the other phosphatidylcholines. The thermograms of the DPPC liposomal bilayers incorporated with the hexane extract of godulbaegi(Ixeris sonchifolia H.) were obtained, and the enthalpy changes and the sizes of cooperative unit of the transition were calculated. The incorporation of the Ixeris sonchifolia H. into the liposomal bilayers effectively reduced the transition temperature at which the transition from gel state to liquid-crystalline state occurs, broadened the thermogram peaks, and reduced the ratio of van't Hoff to calorimetric enthalpies. These results indicate indicate that the godulbaegi extracts (Ixeris sonchifolia H.) have significant effects on the fluidity of biological membrance.

• Journal of Environmental Science International
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• v.12 no.1
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• pp.77-80
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• 2003

The interaction of mastoparan B, a cationic tetradecapeptide amide isolated from the hornet Vespa basalis, with phospholipid bilayers was studied with synthetic mastoparan B and its analogue with Ala instead of hydrophobic 12th amino acid residue in mastoparan B. MP-B and its derivative, [12-Ala]MP-B were synthesized by the solid-phase peptide synthesis method. MP-B and its analogue, [12-Ala]MP-B adopted an unordered structure in buffer solution. In the presence of neutral and acidic liposomes, the peptides took an $\alpha$-helical structure. The two peptides interacted with neutral and acidic lipid bilayers. These results indicated that the hydrophobic face in the amphipathic $\alpha$-helix of MP-B critically affected the biological activity and helical content.

• The Korean Journal of Pharmacology
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• v.26 no.1
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• pp.77-82
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• 1990

The effects of phenothiazine derivatives and calcium on the thermotropic phase transition of bilayers in dipalmitoyl phosphatidylcholine (DPPC) and dipalmitoyl phosphatidic acid (DPPA) liposomes were investigated with differential scanning calorimeter (DSC). Bilayers underwent an abrupt organizational changes at a characteristic temperature when heated. Such temperature-dependent transition was particularly striking and sharp in the bilayers prepared from pure phospholipids. The ability of phenothiazine derivatives to modify the phase transition of phospholipids liposomes was measured by a broadening of the phase transition profile, that transition began to appear at lower temperature than which occurs in untreated liposomes. Calcium ion caused a large upward shift in the transition temperature of DPPC:DPPA (34:66mol%) liposomes. When the liposomes were first incubated with calcium ion followed by phenothiazine derivatives, disappearance of the broad curve centering at $73^{\circ}C$ indicated displacement of calcium ion by phenothiazine derivatives at the anionic site. It is supposed that calcium ion and phenothiazine derivatives might compete with each other on the head group of acidic phospholipid.

• Archives of Pharmacal Research
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• v.11 no.3
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• pp.181-184
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• 1988

The effect of clover saponin on the phase transition of liposomal lipid bilayers of dimyristoyl phosphatidylcholine was investigated with differential scanning calorimetry. The thermograms of the liposomal bilayers incorporated with the clover saponin were obtained, and the enthalpy changes and the sizes of cooperative unit of the transition were calculated. The results showed that incorporation of the clover saponin into the liposomal bilayers effectively reduced the transition temperature at which the transition from solid state to liquid-crystalline state occurs, and broadened the thermogram peaks. It also reduced the size of cooperative unit of the transition. These results indicate that the clover saponin might have significant effect on the fluidity of biological membranes.