• Title/Summary/Keyword: membrane vesicles

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Reconstitution of Membrane Proteins from Corn Seedlings with Phosphatidylcholine (옥수수 유식물 조직에서 분리한 막 단백질과 Phosphatidylcholine의 재조합)

  • 오승은
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.321-323
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    • 1990
  • Membrane proteins isolated from the coleoptile and mesocotyl tissues of corn seedlings were solubilized with Triton X100 and reconstituted with phosphatidylcholine at 2$0^{\circ}C$. The proteoliposomes were incubated and proton uptake into the vesicles was measured with a spectrophotometer. Addition of ATP to the reaction mixture was found to result in an active accumulation of proton into the vesicles. These results indicate that the preparation contains tightly bound phosphatidylcholine vesicles with reconstituted H+ -ATPase from the plant cell membranes.

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Purification and Characterization of ATPase and Phosphatase of Light Membrane Vesicles Isolated from Cucurbita pepo (Cucurbita pepo에서 분리한 Light Membrane Vesicle의 ATPase와 Phosphatase의 정제 및 특성)

  • 오승은
    • Journal of Plant Biology
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    • v.33 no.4
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    • pp.325-332
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    • 1990
  • Light membrane vesicles were isolated from the zucchini hypocotyl by floatation on ficoll density gradients and the proteins were solubilized with Triton X100. Three ATP-hydrolyzing enzymes were partially purified by ion-exchange and gel filtration chromatography and isoelectric focusing. There are plasma membrane-type ATPase whose activity was inhibited by vanadate but not by nitrate, tonoplast-type ATPase which was sensitive to nitrate but insensitive to vanadate and one having a phosphatase activity with a pI value different from that of an acid phosphatase. A fraction was obtained after DEAE-ion-exchange chromatography crossreacting with polyclonal antibodies against Ca2+ -ATPase from human erythrocytes.

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Increase in $Na^+-Ca^{2+}$ Exchange Activity in Sarcolemma Isolated from Mesenteric Arteries of Spontaneously Hypertensive Rats

  • Lee, Shin-Woong;Lee, Jeung-Soo;Park, Young-Joo;Park, In-Sook
    • Archives of Pharmacal Research
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    • v.12 no.2
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    • pp.128-134
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    • 1989
  • $Na^+-Ca^{2+}$ exchange process in sarcolemmal vesicles isolated from mesenteric arteries of Wistar-Kyoto normotensive(WKY) and spontaneously hypertensive rats(SHR) was investigated. The sarcolemmal fractions isolated after homogenization and sucrose density gradient centrifugation were enriched with 5'-nucleotidase and ouabain sensitive, $K^+-dependent$ phosphatase activities. When the vesicles were loaded with $Na^+$, a time dependent $Ca^{2+}$ uptake was observed. However, very little $Ca^{2+}$ uptake was observed when the vesicles were loaded with $K^+$, or $Ca^{2+}$ uptake of the $Na^+-loaded$ vesicles was carried out in high sodium medium so that there was no sodium gradient. When the vesicles loaded with $Ca^{2+}$ by $Na^+-Ca^{2+}$ exchange were diluted into potassium medium containing EGTA, $Ca^{2+}$ was rapidly released from the vesicles. $Na^+-dependent\;Ca^{2+}$ uptake was increased in SHR compared to WKY, but passive efflux of preaccumulated $Ca^{2+}$ from the vesicles was decreased in SHR. The data indicate that the membrane vesicles of rat mesenteric arteries exhibit $Na^+-Ca^{2+}$ exchange activity. It is also suggested that changes of this process in vascular smooth muscle cell membrane of SHR may be involved in higher intracellular $Ca^{2+}$ concentration and higher basal tone in SHR.

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Effects of D-Fructose on the Uptake of Iron by the Intestinal Brush-Border Membrane Vesicles from Rats.

  • Kim, Ok-Seon;Lee, Yong-Bok;Oh, In-Joon;Koh, Ik-Bae;Lee, Yeong-Woo
    • Journal of Pharmaceutical Investigation
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    • v.24 no.3
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    • pp.11-18
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    • 1994
  • We have studied the iron uptake by the purified brush-border membrane vesicles (BBMVs) to determine the effect of fructose on the absorption of iron. BBMVs were prepared by the modified calcium precipitation method, The degree of purification was routinely assessed by the marker enzyme, alkaline phosphatase, and the functional integrity was tested by $D-[1-^3H]glucose$ uptake. The appearance of membrane vesicles was shown by transmission electron microscopy (TEM). The uptakes of complexes of labeled iron $[^{59}Fe]$ with fructose and ascorbate were measured with a rapid filtration technique, The uptake rate and pattern of the two iron-complexes, Fe(III)-fructose and Fe(III)-ascorbate, were also observed. A typical overshooting uptake of D-glucose was observed with peak value of $2{\sim}3$ times higher concentration than that at equilibrium. This result was similar to other studies with BBMVs. TEM showed that the size of BBMVs was uniform and we can hardly find any contaminants, Fe(III)-fructose has the higher value of $V_{max}$ and the lower value of Km than those of Fe(III)-ascorbate, respectively. It may be concluded that D-fructose is more effective in promoting the iron absorption than ascorbate.

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Formation of Dictyosome and Spherosome in Endosperm Cells of Panax ginseng during seed Formation (인삼(人蔘)(Panax ginseng)의 종자형성(種子形成)에 따른 배유세포(胚乳細胞)의 딕티오좀 및 Spherosome 형성)

  • Yu, Seong-Cheol;Kim, Woo-Kap
    • Applied Microscopy
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    • v.21 no.2
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    • pp.117-125
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    • 1991
  • This study has been carried out to investigate the development of dictyosome, and roles of dictyosome about the formation of spherosome in the endosperm cell during seed formation of Panax ginseng with electron microscope. The result is as follows; In the endosperm cells of early stage during seed formation of Panax ginseng, plastid, mitochondria, endoplasmic reticulum, dictyosome and ribosomes are evenly distributed in cytoplasm. Electron lucent vesicles derived from dictyosome are observed in endosperm cells. Vesicles that contain low electron density are derived from forming face of dictyosome and releases into the cytosol. This vesicles formed multi vesicular body or fused with the plasma membrane. The spherical spherosomes are formed from dictyosome containing the lipid materials of even electron density and are gradually increased in size and number. Dictyosome is located in between vacuole and spherosome and it's cisternae form a semicircle and a circle. Some membrane of the protein body that accumulate the storage protein are originate from the spherical vacuole which interfused between vesicles and vacuoles derived from dictyosome.

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Simple Analysis for Interaction between Nanoparticles and Dye-Containing Vesicles as a Biomimetic Cell-Membrane

  • Shin, Sohyang;Umh, Ha Nee;Kim, Younghun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.1
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    • pp.231-236
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    • 2013
  • Some cytotoxicity studies for the interpretation of the interaction between nanoparticles and cells are non-mechanistic and time-consuming. Therefore, non-biological screening methods, which are faster and simpler than in-vivo and in-vitro methods, are required as alternatives to current cytotoxicity tests. Here, we proposed a simple screening method for the analysis of the interaction between several AgNPs (bare-, citrate-, and polyvinylpyrrolidone-coating) and dye-containing vesicles acting as a biomimetic cell-membrane. The interaction between AgNPs and vesicles could be evaluated readily by UV-vis spectra. Absorbance deviation in UV-vis spectra revealed a large attraction between neighboring particles and vesicles. This was confirmed by (Derjagin, Landau, Verwey, and Overbeek) theory and DMF (dark-field microscopy) analysis. This proposed method might be useful for analyzing the cytotoxicity of nanoparticles with cell-membranes instead of in vitro or in vivo cytotoxicity tests.

Microbe-derived extracellular vesicles as a smart drug delivery system

  • Yang, Jinho;Kim, Eun Kyoung;McDowell, Andrea;Kim, Yoon-Keun
    • Translational and Clinical Pharmacology
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    • v.26 no.3
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    • pp.103-110
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    • 2018
  • The human microbiome is known to play an essential role in influencing host health. Extracellular vesicles (EVs) have also been reported to act on a variety of signaling pathways, distally transport cellular components such as proteins, lipids, and nucleic acid, and have immunomodulatory effects. Here we shall review the current understanding of the intersectionality of the human microbiome and EVs in the emerging field of microbiota-derived EVs and their pharmacological potential. Microbes secrete several classes of EVs: outer membrane vesicles (OMVs), membrane vesicles (MVs), and apoptotic bodies. EV biogenesis is unique to each cell and regulated by sophisticated signaling pathways. EVs are primarily composed of lipids, proteins, nucleic acids, and recent evidence suggests they may also carry metabolites. These components interact with host cells and control various cellular processes by transferring their constituents. The pharmacological potential of microbiome-derived EVs as vaccine candidates, biomarkers, and a smart drug delivery system is a promising area of future research. Therefore, it is necessary to elucidate in detail the mechanisms of microbiome-derived EV action in host health in a multi-disciplinary manner.

Effect of Temperature on Dicarboxylate Transport in Plasma Membrane Vesicles of Rabbit Proximal Tubule

  • Han, Kyung-Moon;Kim, Young-Hee;Woo, Jae-Suk;Kim, Yong-Keun
    • The Korean Journal of Physiology
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    • v.27 no.2
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    • pp.227-232
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    • 1993
  • The temperature dependence of $Na^+-dependent$ succinate uptake was studied in brush border (BBMV) and basolateral (BLMV) membrane vesicles isolated from the rabbit kidney cortex. The succinate uptake was markedly altered by temperature in a similar fashion in both membranes. The temperature dependence was characterized by a nonlinear Arrhenius plot with a break point at 22 and $25^{\circ}C$ for BBMV and BLMV, respectively. The activation energy was 3.91 and 17.09 kcal/mole at above and below the break point respectively, far BBMV; 2.65 and 14.05 kcal/mole, respectively, for BLMV. When temperature increased f개m 20 to $35^{\circ}C$, the Vmax of succinate transport increased from $3.49{\pm}0.11\;to\;5.90{\pm}0.86\;nmole/mg/5\;sec$ for BBMV and from $2.86{\pm}0.25\;to\;3.63{\pm}0.32\;nmole/mg/5\;sec$ for BLMV, with no change in Km in both membranes. These results suggest that renal dicarboxylate transport is similarly sensitive to a change in membrane physical state in BBMV and BLMV.

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Outer Membrane Vesicles Derived from Salmonella Enteritidis Protect against the Virulent Wild-Type Strain Infection in a Mouse Model

  • Liu, Qiong;Yi, Jie;Liang, Kang;Zhang, Xiangmin;Liu, Qing
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1519-1528
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    • 2017
  • Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

Effects of Ethanol on $Na^+-dependent$ Solute Uptake in Rabbit Renal Brush-Border Membrane Vesicles

  • Kim, Yong-Keun;Ko, Sun-Hee;Woo, Jae-Suk;Jung, Jin-Sup;Lee, Sang-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.2
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    • pp.191-198
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    • 1999
  • This study was undertaken to examine the effect of ethanol on $Na^+-dependent$ transport systems (glucose, phosphate, and dicarboxylate) in renal brush-border membrane vesicles (BBMV). Ethanol inhibited $Na^+-dependent$ uptakes of glucose, phosphate, and succinate in a dose-dependent manner, but not the uptakes of $Na^+-dependent.$ The $H^+/TEA$ antiport was reduced by 8% ethanol. Kinetic analysis showed that ethanol caused a decrease in $V_{max}$ of three transport systems, leaving $K_m$ values unchanged. Ethanol decreased phlorizin binding, which was closely correlated with the decrease in $V_{max}$ of $Na^+-glucose$ uptake. These results indicate that ethanol inhibits $Na^+-dependent$ uptakes of glucose, phosphate, and dicaboxylate and that the reduction in $V_{max}$ of $Na^+-glucose$ uptake is caused by a decrease in the number of active carrier proteins in the membrane.

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