• BMB Reports
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• 제44권10호
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• pp.674-679
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• 2011

Bacillus sphaericus produces mosquito-larvicidal binary toxin composed of BinA and BinB. While BinB is expected to bind to a specific receptor on the cell membrane, BinA interacts to BinB or BinB receptor complex and translocates into the cytosol to exert its activity via unknown mechanism. To investigate functional roles of aromatic cluster in BinA, amino acids at positions Y213, Y214, Y215, W222 and W226 were substituted by leucine. All mutant proteins were highly produced and their secondary structures were not affected by these substitutions. All mutants are able to insert into lipid monolayers as observed by Langmuir-Blodgett trough and could permeabilize the liposomes in a similar manner as the wild type. However, mosquito-larvicidal activity was abolished for W222L and W226L mutants suggesting that tryptophan residues at both positions play an important role in the toxicity of BinA, possibly involved in the cytopathological process after toxin entry into the cells.

• Journal of Pharmaceutical Investigation
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• 제21권3호
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• pp.161-170
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• 1991

The storage stabilities of immunoliposomes incorporated with variable amounts of ganglioside $G_{M1}$ were investigated as a function of time. temperature. and composition by observing absorbance of visible light and calcein release. In the column chromatographe, the layer of unsaturated PE(dioleoylphosphatidylethanolamine : DOPE). unable to form stable liposomes at physiological temperature and pH, were formed when palmitoyl-immunoglobulin G(IgG) $(2.5{\times}10^{-4}\;mol/DOPE\;mol)$ added. The incorporation of ganglioside $G_{M1}$ into immunoliposome. enhanced the stabilities of bilayers during the extended period of storage. The turbidities of immunoliposomes coated with ganglioside $G_{M1}$ exhibited the maximum near 20 mol% $G_{M1}/DOPE$ mol. probably because of the disturbance of the bilayer characteristics, i.e., layer transition or reorientation of interaction sites. At low temperature. the higher stability was achieved than at elevated temperatures. After one week of storage. the redispersed liposomal solutions at lower temperatures maintained the original elution patterns in chromatography but broader distribution at elevated temperatures. During the storage, it is suggested the aggregation is the more dominant phenomena for liposomes kept at $5^{\circ}C$ than the fusion. while he fusion is at elevated temperatures.

• Archives of Pharmacal Research
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• 제18권2호
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• pp.84-89
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• 1995

The complexation and physical characteristics of egg phosphatidylcholine (PC) liposome containing amphotericin B(AmB) were investigated through circular dichrosim(CD) spectra, the size distribution, the turbidity change, and the calcein release. CD spectra of AmB-containing egg PC mxture exhibited a positive peak around 330 nm indicative of complexation of AmB and four negative peaks. The positive peak increased up to $2.2{\;}millidegree/{\mu}g$ AmB as AmB contents increased up to 12% (w/w), suggesting that AmB-phospholipid complexation was promoted by the antibiotics. The effective diameter of liposomesa by dynamic light scattering decreased from 450 nm to 220 nm as the amount of AmB in liposomes increased from o to 30% (w/w). The complexation may be responsible for the reduction in size. On the other hand, at around 1 mN deoxycholate (DOC), the reltive turbidities of 5 and 10% (w/w) AmB-containing liposome suspension were less than 1 probably due to the soblubilization of the complex, while those of pure PC liposome suspension were larger than 1 at the same concentration. Deoxycholate-induced release of liposomes, indicating the intercalation of the drug into the bilayers. Therefore, it is concluded that in AmB/eggPC/water system, AmB-phospholipid complexcoexists with AmB-containing liposomes.