• 한국해양바이오학회지
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• 제2권2호
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• pp.123-125
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• 2007

Vibrio anguillarum is a gram-negative bacterium that causes vibriosis in approximately 80 different fish species. V. anguillarum produces several exotoxins are correlated with the pathogenesis of vibriosis. This study is focused on the composition of the outer membrane vesicle. Most of gram-negative bacteria produce outer membrane vesicle (OMV) during cell growth. OMV was formed from the outer membrane surface of cell and than released to extracellular environment. OMV consists of outer membrane lipids, outer membrane protein (OMP), LPS, and soluble periplasmic components. Also, they contain toxins, adhesions, and immunomodulatory. Many gram-negative bacteria were studied out forming OMV. In Vibrio sp., formation of OMV by electron microscopy has been reported from V. cholerae and V. parahaemolyticus. In present study, we isolated OMV from V. anguillarum and OMV protein was separated by SDS-PAGE. Magor band was sliced and analyzed by MALDI-TOF. The major protein band of 38kDa was identified as OmpU by MALDI-TOF MS analysis.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1343-1347
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• 2016

Outer membrane vesicles (OMVs) are spherical nanostructures that are ubiquitously shed from gram-negative bacteria both in vitro and in vivo. Recent findings revealed that OMVs, which contain diverse components derived from the parent bacterium, play an important role in communication with neighboring bacteria and the environment. Furthermore, nanoscale proteoliposomes decorated with pathogen-associated molecules attract considerable attention as a non-replicative carrier for vaccines and drug materials. This review introduces recent advances in OMV biogenesis and discusses the roles of OMVs in the context of bacterial communication and virulence regulation. It also describes the remarkable accomplishments in OMV engineering for diverse therapeutic applications.

• 대한약리학회지
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• 제28권1호
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• pp.103-114
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• 1992

한국산 2년생 소의 신선한 대뇌피질로 부터 synaptosomal plasma membrane vesicles (SPMV)를 분리한 후 이 SPMV로 부터 추출한 총지질 (SPMVTL) 및 총인지질 (SPMVPL)로서 인공세포막을 제제하였다. SPMVTL 및 SPMVPL의 outer monolayer에 trinitrophenyl groups로 공유결합시킴으로써 형광 probe 1,6-diphenyl-1,3,5-hexatriene (DPH)의 outer monolayer 형광을 선택적으로 소광케한 후 SPMVTL 및 SPMVPL의 inner와 outer monolayer 사이의 비대칭적 유동성 존재 유무를 확인하였던 바 inner monolayer에 비하여 outer monolayer가 유동성이 크다는 것을 확인하였으며 SPMVTL에 비하여 SPMVPL의 유동성이 높았다. 뿐만 아니라 barbiturates는 SPMVTL의 유동성에 대하여는 유의성이 있을 만큼의 증감 효과를 나타내지 않았고 SPMVPL의 유동성은 감소시키되 주로 outer monolayer의 유동성을 감소시킨다는 것을 확인하였으며 barbiturates 종류에 따른 SPMVPL의 총유동성 감소의 크기는 pentobarbital > hexobarbital > amobarbital > phenobarbital의 순이었다.

• 대한약리학회지
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• 제28권2호
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• pp.191-199
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• 1992

한국산 2년생 소의 대뇌피질로부터 synaptosomal plasma membrane vesicles(SPMV)를 분리한 후 이 SPMV로부터 추출한 총지질로서 인공세포막(SPMVTL)을 제제하였다. SPMVTL의 outer monolayer에 trinitrophenyl group을 공유결합시킴으로써 SPMVTL이중층에 분포된 형광 probe 1,6-diphenyl-1,3,5-hexatriene(DPH)중 outer monolayer에 분포된 DPH형광만을 소광케 하였다. 형광분석기를 통하여 SPMVTL의 inner monolayer에 비하여 outer monolayer의 유동성이 크다는 것을 확인하였다. n-Alkanols는 SPMVTL지질 이중층(inner+outer monolayers)의 회전확산운동을 증가시키되 탄소수 두개가 증가될 때마다 그 효력이 약10배 가량 증가된다는 것을 알았다. 그러나 1-decanol의 경우에는 1-nonanol에 비하여 그 효력이 낮아지는 소위 cut-off현상이 나타났다. 뿐만 아니라 n-alkanols는 SPMVTL의 inner monolayer에 비하여 outer monolayer의 회전확산운동을 주로 증가시켰다. 하지만 n-alkanols의 탄소수가 증가됨에 따라 outer monolayer에 대한 선택적인 작용이 감소된다는 것도 확인되었다.

• The Korean Journal of Pain
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• 제6권2호
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• pp.237-246
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• 1993

세포막에서 마취제의 작용점을 규명하기 위하여, 마취제의 많은 부분을 차지하는 n-Alkanol을 이용하여, 소의 synaptosomal plasma membrane vesicles(SPMV)에서 n-Alkanol의 침투 정도를 형광 probe를 이용한 형광소광법을 통하여 검색하였다. n-Alkanols는 SPMV 외부 단층(outer monolayer)의 표면에 주로 분포하되 그 탄소수에 비례하여 소수성 부위에 분포되는 양이 증가되는 경향을 나타내었다(1-decanol은 제외). Methanol, Ethanol, 1-propanol, 1-butanol, 1-pentano, 1-hexanol, 1-heptanol, 1-octanol, 1-nonanol 및 1-decanil은 SPMV 외부 단층의 표면(친수성 부위)에 분포되는 것에 비하여 각각 949, 416.8, 214.8, 90.3, 53.7, 15.20, 6.80, 2.00, 1.03 및 2.40 배가 된다는 것을 확인하였다. 1-decanol은 $C_{10}$인데도 불구하고 $C_8$인 1-octanol에 비하여 적은 양이 소수성 부위에 침투 분포한다는 것이 확인되었다.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1271-1279
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• 2009

To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

• 한국미생물·생명공학회지
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• 제27권6호
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• pp.440-445
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• 1999

In order to investigate structure-antibiotic activity relationships of brevinin-1 and thanatin containing Rana box composed of basic loop formed by disulfide bridge in their arboxy terminus, thanatin, brevinin 1 and their analogues (T-B1, T-B2 and B-T) in which their Rana box sequence exchanged was designed and synthesized by the solid phase method using Fmoc-chemistry. The basic sequence of Rana box of thanatin had more significant effect on both antibacterial and antifungal activity than that of brevinin 1. The tail sequence (QRM) of thanatin was found to be important in its antibacterial and antifungal activity. Rana box sequence of brevinin-1 did not have a significant effect on its antitumor and phospholipid vesicle-aggregating activities. Brevinin-1 showed stronger $\alpha$-helical structure in the membrane-mimicking environment such as SDS micelle than thanatin. A remarkable increase in a-helicity of bervinin-1 plays more important role in antibiotic activity than that of thanatin. Furthermore, antibacterial activity of thanatin against E. coli resulted from the disruptive effect against the outer cell membrane of E. coli.

• Journal of Oral Medicine and Pain
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• 제24권4호
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• pp.361-374
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• 1999

F. nucleatum is a gram-negative obligate anaerobe which is the principal and most frequent cause of gingival inflammation and is the predominant pathogen isolated in subsequent periodontal breakdown. It is also one of the most numerous bacteria found in subgingival plaque samples from healthy sites; its numbers are about 10-fold greater in plaque from periodontally diseased sites. The purpose of this study is to examine the effects of outer membrane(OM), outer membrane vesicle(OMV), and lipopolysaccharide(LPS) from F. nucleatum ATCC 25586 strain on the growth of human gingival fibroblasts and HOS 941 cells, and on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes. For the examination of cytotoxic effects, $TNF-{\alpha}$ production and $TNF-{\alpha}$ mRNA expression, the MTT assay, the ELISA and the RT-PCR were performed, respectively. All extracts of F. nucleatum tested were cytotoxic to both of human gingival fibroblasts and HOS 941 cells, and the significant difference of cytotoxic activity among the extracts was not observed. In the effects of these extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression of mouse splenocytes, all extracts of F. nucleatum tested also stimulated the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression, but the effects of the OM extracts on the $TNF-{\alpha}$ production / $TNF-{\alpha}$ mRNA expression were higher than those of the OMV and the LPS extracts. The pattern of the $TNF-{\alpha}$ mRNA expression was similar to that of the $TNF-{\alpha}$ production. These results indicate that F. nucleatum seems to contribute to the pathogenesis of periodontal diseases at least by its cytotoxicity, directly and its $TNF-{\alpha}$ production, indirectly.

• The Korean Journal of Physiology and Pharmacology
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• 제3권5호
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• pp.513-519
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• 1999

Direct exposure of renal tubular brush-border membranes (BBM) to free cadmium (Cd) causes a reduction in phosphate (Pi) transport capacity. Biochemical mechanism of this reduction was investigated in the present study. Renal proximal tubular brush-border membrane vesicles (BBMV) were isolated from rabbit kidney outer cortex by Mg precipitation method. Vesicles were exposed to $50{\sim}200\;{\mu}M\;CdCl_2$ for 30 min, then the phosphate transporter activity was determined. The range of Cd concentration employed in this study was comparable to that of the unbound Cd documented in renal cortical tissues of Cd-exposed animals at the time of onset of renal dysfunction. The rate of sodium-dependent phosphate transport $(Na^+-Pi\;cotransport)$ by BBMV was determined by $^{32}P-Iabeled$ inorganic phosphate uptake, and the number of $Na^+-Pi$ cotransporters in the BBM was assessed by Pi-protectable $^{14}C-labeled$ phosphonoformic acid $([^{14}C]PFA)$ binding. The exposure of BBMV to Cd decreased the $Na^+-Pi$ cotransport activity in proportion to the Cd concentration in the preincubation medium, but it showed no apparent effect on the Pi-protectable PFA binding. These results indicate that an interaction of renal BBM with free Cd induces a reduction in $Na^+-Pi$ cotransport activity without altering the carrier density in the membrane. This, in turn, suggest that the suppression of phosphate transport capacity $(V_{max})$ observed in Cd-treated renal BBM is due to a reduction in $Na^+-Pi$ translocation by existing carriers, possibly by Cd-induced fall in membrane fluidity.

• 대한약리학회지
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• 제29권1호
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• pp.157-163
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• 1993

n-Alkanols의 분자적 약리작용기전 탐구에 기초자료를 제공하기 위하여 소의 신선한 대뇌피질로부터 분리한 synaptosomal plasma membrane vesicles (SPMV) 지질 이중층의 측방확산운동에 미치는 n-alkanols의 영향을 형광 probe법으로 검색하였다. n-Alkanols는 SPMV 지질 이중층의 측방확산운동 범위와 속도를 농도 의존적으로 증가시켰고 1-nonanol까지는 탄소수가 두개 증가될 때마다 그 효력은 약 10배 가량 증가되었으나 탄소수 10개인 1-decanol의 효력은 오히려 감소되는 경향을 나타내었다.