• BMB Reports
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• v.29 no.2
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• pp.169-174
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• 1996

Yeast cytochrome c (cyt c) was modified at cysteine-102 with a thiol-specific spin label and its interaction with liposomes containing acidic phospholipids was studied by electron paramagnetic resonance (EPR) spectroscopy. Association of cyt c with liposomes resulted in a significant reduction in the mobility of the spin label and a fraction of cyt c even seemed to be immobilized. Based on a large spectral change upon binding and the proximity of the spin-label to lysine-86 and -87, we propose these two residues to be the potential binding site at neutral pH. The interaction is electrostatic in nature because the spectral changes were reversed by addition of anions. Dissociation of the bound cyt c by anions, however, became less effective as the lipid/protein ratio increased. This suggests a repulsive lateral interaction among the bound cyt c. Unlabeled cyt c molecules added to preformed cyt c-liposome complex displaced the bound (spin labeled) cyt c and the process was competitive and reversible.

• Applied Chemistry for Engineering
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• v.28 no.2
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• pp.177-185
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• 2017

In the present work, the interaction of fatty acid with vesicle membrane of phospholipids was investigated using 3 different kinds of fatty acids such as stearic acid (SA), oleic acid (OA) and linoleic acid (LA). Basically, the same trend has been found in 3 fatty acid systems. The addition of fatty acid produced a close packing of liposome due to the penetration of fatty acid molecules into liposome vesicles, which resulted in a decrease in size and an increase in zeta potential of liposome. However, excessive addition of fatty acid produced a transition from liposomes to aggregates of lipid particles having polymorphic structure. The membrane fluidity, characterized by measuring membrane deformability and fluorescence anisotropy ratio of liposomes, was in good agreement with measurement results of transmission electron microscopy (TEM) and particle size. The minimum size and closest packing of liposome with SA, OA and LA were found when the molar ratios of fatty acid to lecithin were 0.70, 0.50, and 0.25 respectively.

• 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 Yeungnam Medical Science
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• v.3 no.1
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• pp.41-48
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• 1986

Band 3, the predominent 95,000 dalton anion transport protein, is the major intrinsic glycoprotein of the human erythrocyte membrane. This anion carrier exists as a dimer and binds the cytoskeletons such as spectrin, ankyrin and actin. And the liposomes are vesicular structures which form spontaneouly upon hydration of phospholipids. These artificial lipid vesicles have been investigated as model of the biological membranes and as a mean of improving the delivery of nucleic acids, drugs, proteins and biological substances to specific target tissues and cells. In this study, we were purified Band 3 from the human erythrocyte membrane(ghost) was prepared by hemolysis of intact human erythrocyte with weak alkali-hypotonic solution. Band 6 was removed from ghost by extracting with solution of an ionic strength of 0.15. Band 3 and Band 4 were solubilized selectively by extracting Band 6-depleted ghosts with Triton X-100 under nondenaturing conditions. Band 3 was then purified from Triton X-100 extract treated with p-chloromercuribenzoate by sucrose density gradient ultracentrifugation. This purified Band 3 was incorporated into liposomes prepared by reverse-phase evaporation. Phosphatidyl L-serine and cholesterol(1 : 1 molar ratio) were dissolved in chloroform and then chloroform was removed by rotatory evaporation under reduced pressure. Band 3 solution without Triton X-100 was introduced into a mixture of lipids and diethylether. Diethylether was subsequently removed by evaporation. This purified Band 3 and its incorporation into liposomes were confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

• International Journal of Oral Biology
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• v.42 no.4
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• pp.175-181
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• 2017

The aim of this study was to provide a basis for the molecular mechanism underlying the pharmacological action of ethanol. We studied the effects of 1-propanol on the location of n-(9-anthroyloxy)palmitic acid or stearic acid (n-AS) within the phospholipids of synaptosomal plasma membrane vesicles (SPMV). The SPMV were isolated from the bovine cerebral cortex and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL). 1-Propanol increased the rotational mobility of inner hydrocarbons, while decreasing the mobility of membrane interface, in native and model membranes. The degree of rotational mobility varied with the number of carbon atoms at positions 16, 12, 9, 6 and 2 in the aliphatic chain of phospholipids in the neuronal and model membranes. The sensitivity of increasing or decreasing rotational mobility of hydrocarbon interior or surface by 1-propanol varied with the neuronal and model membranes in the following order: SPMV, SPMVPL and SPMVTL.

• BMB Reports
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• v.28 no.2
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• pp.129-137
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• 1995

The ryanodine-receptor $Ca^{2+}$ release channel protein in the sarcoplasmic reticulum membrane of rabbit skeletal muscle plays an important role in muscle exitation-contraction (E-C) coupling. Various types of detergents were tested, including Chaps, cholate, octylglucoside, Zwittergents, Mega-9, Lubrol PX, and Triton X-100 for solubilization of this protein. Among these, Chaps and Triton X-100 were found to optionally solubilize the channel complex. Optimum conditions for this solubilization were pH 7.4 with a salt concentration of 1 M. The addition of phospholipid in the solubilization step helped in stabilizing the protein. The purification of the receptor was performed using sucrose density gradient centrifugation. Various methods [dilution, freeze-thaw, adsorption (Biobeads), and dialysis] were investigated to incorporate the Chaps-solubilized and purified $Ca^{2+}$ release channel protein into liposomes made from different types of phospholipids. Of these, a combined method consisting of a dialysis, freeze-thaw and sonication steps yielded the best results. Reconstituted vesicles produced by this method with 95% phosphatidylcholine (from soybean extract) had good function.

• Polymer(Korea)
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• v.38 no.6
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• pp.694-701
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• 2014

In this study, Polygomun aviculare L. (P. aviculare L.) extract loaded elastic liposomes (ELPs) were investigated to enhance the transdermal delivery of P. aviculare L. extract composed of various flavonoids. ELPs were composed of egg phospholipids (PC) and edge activator ($Tego^{(R)}$ care 450) and the physical properties and in vitro permeation studies of ELPs were performed. Particle size ranged from 148.1 to 262.2 nm and deformability index was recorded as 11.5~25.4. Loading efficiency was from 53.1 to 66.3%. In vitro skin permeation studies using Franz diffusion cell demonstrated that ELP-4 having ratio of 85:15 for PC to $Tego^{(R)}$ care 450 exhibited the higher skin permeability than ELP-1, the general liposome without $Tego^{(R)}$ care 450. It was visually seen by fluorescence image restoration microscopy. The findings suggest that ELP-4 selected as the optimal formulation could be used as useful formulation for transdermal delivery of the extract.

• Polymer(Korea)
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• v.36 no.6
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• pp.705-711
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• 2012

In this study, the elastic liposome consisted of egg phospholipids and edge activator ($Tego^{(R)}$ care 450) was prepared in order to supplement the defect of the conventional liposome. We prepared elastic liposome containing quercetin, known as natural antioxidant, and evaluated the vesicles size, elasticity, loading efficiency, stability, and in vitro skin permeation. The mean diameter of quercetin loaded elastic liposome formulations ranged between 208.2~303.4 nm and loading efficiency was observed 64.1~87.5%. The highest loading efficiency (87.5%) and deformability (28.3) were observed at the optimal ratio of 90 : 10 (egg phospholipids : $Tego^{(R)}$ care 450) among 0.1% quercetin loaded elastic liposome formulations. The elastic liposome formulation was selected for further transdermal permeation study. The elastic liposome ($129.9{\mu}g/cm^2$) exhibited more skin permeability than general liposome ($114.8{\mu}g/cm^2$) and 1,3-butylene glycol ($75.1{\mu}g/cm^2$) solution. This results suggest that the elastic liposome formulation using $Tego^{(R)}$ care 450 as a major edge activator could be useful for the delivery of active ingredient through the skin transdermal.

• Applied Biological Chemistry
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• v.38 no.4
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• pp.320-324
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• 1995

Solubilization of microsomal ${\Delta}^{5}-3{\beta}$-hydroxy steroid acyl transfearse(${\Delta}^{5}-3{\beta}$-OH-SAT) of rat brain and its reconstitution into liposomes were investigated. Among the detergents utilized for the solubilization, deoxycholic acid was superior to Tween 80 or Triton X-100 for the reconstituted activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. The enzyme activity was shown to be affected by the nature of phospholipids used for the preparation of the liposome. Phosphatidylcholines from egg yolk and soybean showed the highest activity of ${\Delta}^{5}-3{\beta}$-OH-SAT and phosphatidylethanolamine came next. However phosphatidylserine and phosphatidic acid showed a lower activity than those obtained before the reconstitution. This study suggests that the presence of quaternary ammonium salt or amine group in the phospholipids stimulates the activity of ${\Delta}^{5}-3{\beta}$-OH-SAT. However the presence of a carboxylic group or the absence of the amine group may have an inhibitory effect on the ${\Delta}^{5}-3{\beta}$-OH SAT.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.38 no.2
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• pp.147-154
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• 2012

In this study, we prepared elastic liposome containing rutin, known as antioxidants, and evaluated the physical characterization and enhanced skin permeation effect. The elastic liposome was prepared using the different ratios of egg phospholipids and $Tego^{(R)}$ care 450. The mean diameter of rutin loaded elastic liposomes formulations ranged between 205.7 ~ 298.0 nm and deforability 20.9 ~ 42.5, The loading efficiency was observed to be 52.0 ~ 71.0 %. The highest loading efficiency (71.0 %) and deformability (42.5) were observed at the optimal ratio of 85 : 15 (egg phospholipids : $Tego^{(R)}$ care 450) in the 0.1 % rutin loaded elastic liposome formulations. The elastic liposome formulation was selected for further transdermal permeation study. The elastic liposome(129.9 ${\mu}g/cm^2$) exhibited a significantly higher skin permeation compared with general liposome (98.0 ${\mu}g/cm^2$) and 1,3-butylene glycol (76.3 ${\mu}g/cm^2$) solution. These results suggest that the elastic liposome formulation using $Tego^{(R)}$ care 450 as a major edge activator could be useful for the delivery of active ingredient through the skin barrier.