• The Korean Journal of Physiology
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• 제22권2호
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• pp.307-318
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• 1988

가토 신장 피질에서 Percoll density gradient방법으로 분리한 basolateral membrane vesicle (BLMV)에서 rapid filtration technique을 이용하여 succinate의 이동 특성을 관찰하였다. $Na^+$은 succinate의 이동을 증가시켜 "overshoot"현상을 보였으며 이러한 효과는 $K^+,{\;}Li^+,{\;}Rb^+,{\;}choline$과 같은 다른 양이온들에 의해 나타나지 않았다. $Na^+$농도변화에 따른 succinate의 이동율은 sigmoid모양을 보였고, $Na^+$에 대한 Hill coefficient는 2.0이었다. soccinate의 이동은 vesicle 내부가 음전압일 때 더욱 증가되었다. BLMV에서 succinate이동은 용액내 pH변화에 따라 영향을 받았으나 brush border membrane vesicle (BBMV)에서는 영향을 받지 않았다. 동력학적 분석결과 succinate의 Km값은 $15.5{\pm}0.94{\;}{\mu}M$이었고 Vmax는 $16.22{\pm}0.25{\;}n{\;}mole/mg{\;}protein/min$이었다. succinate의 이동은 $4{\sim}5$탄소를 가진 dicarboxylate들에 의해 강력하게 억제되었으나 monocarboxylate나 다른 유기음이온들에 의해 영향을 적게 받거나 받지 않았다. succinate의 이동은 DIDS, SITS, furosemide와 같은 음이온 이동 억제제와 harmaline과 같은 $Na^+$ 이동 억제제에 의해 억제되었다. 이들 결과들은 BLMV에서 succinate는 $Na^+$에 의존하여 이동하며 다른 Krebs cycle중간 산물들과 동일한 운반기전을 이용함을 가르킨다. 또한 BLMV에서 succinate의 이동은 그 기질특이성에 있어서 다른 연구자에 의해 보고된 BBMV에서 이동특성과 유사함을 보였다.

• 한국식품위생안전성학회지
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• 제8권4호
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• pp.189-193
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• 1993

Cisplatin is useful for various cancers including advanced testicular and ovarian cancers. However, clinical use of cisplatin has been limited due to its dose-related neplrotoxicity. Transport studies across the membrane vesicles were performed to study the cisplatin nephrotoxicity. In these experiments, after cisplatin was administered to adult male New Zealand White rabbits, basolateral membrane (BLM) vesicles were prepared from the renal cortex. Para-aminohippurate (PAH) uptakes through BLM vesicles were measured to examine the interactioln of cisplatin on the transports of the substrates. As results of the uptake experiments using the vesicle systems, cisplatin had little effects on PAH transport through BLM vesicle. In conclusion, cisplatin did not cause the damage of basolateral membranes.

• Genomics & Informatics
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• 제19권4호
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• pp.39.1-39.8
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• 2021

Tamoxifen (TAM) is an anticancer drug used to treat estrogen receptor (ER)-positive breast cancer. However, its ER-independent cytotoxic and antifungal activities have prompted debates on its mechanism of action. To achieve a better understanding of the ER-independent antifungal action mechanisms of TAM, we systematically identified TAM-sensitive genes through microarray screening of the heterozygous gene deletion library in fission yeast (Schizosaccharomyces pombe). Secondary confirmation was followed by a spotting assay, finally yielding 13 TAM-sensitive genes under the drug-induced haploinsufficient condition. For these 13 TAM-sensitive genes, we conducted a comparative analysis of their Gene Ontology (GO) 'biological process' terms identified from other genome-wide screenings of the budding yeast deletion library and the MCF7 breast cancer cell line. Several TAM-sensitive genes overlapped between the yeast strains and MCF7 in GO terms including 'cell cycle' (cdc2, rik1, pas1, and leo1), 'signaling' (sck2, oga1, and cki3), and 'vesicle-mediated transport' (SPCC126.08c, vps54, sec72, and tvp15), suggesting their roles in the ER-independent cytotoxic effects of TAM. We recently reported that the cki3 gene with the 'signaling' GO term was related to the ER-independent antifungal action mechanisms of TAM in yeast. In this study, we report that haploinsufficiency of the essential vps54 gene, which encodes the GARP complex subunit, significantly aggravated TAM sensitivity and led to an enlarged vesicle structure in comparison with the SP286 control strain. These results strongly suggest that the vesicle-mediated transport process might be another action mechanism of the ER-independent antifungal or cytotoxic effects of TAM.

• Molecules and Cells
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• 제43권4호
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• pp.313-322
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• 2020

Eukaryotes transport biomolecules between intracellular organelles and between cells and the environment via vesicle trafficking. Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNARE proteins) play pivotal roles in vesicle and membrane trafficking. These proteins are categorized as Qa, Qb, Qc, and R SNAREs and form a complex that induces vesicle fusion for targeting of vesicle cargos. As the core components of the SNARE complex, the SNAP25 Qbc SNAREs perform various functions related to cellular homeostasis. The Arabidopsis thaliana SNAP25 homolog AtSNAP33 interacts with Qa and R SNAREs and plays a key role in cytokinesis and in triggering innate immune responses. However, other Arabidopsis SNAP25 homologs, such as AtSNAP29 and AtSNAP30, are not well studied; this includes their localization, interactions, structures, and functions. Here, we discuss three biological functions of plant SNAP25 orthologs in the context of AtSNAP33 and highlight recent findings on SNAP25 orthologs in various plants. We propose future directions for determining the roles of the less well-characterized AtSNAP29 and AtSNAP30 proteins.

• Bulletin of the Korean Chemical Society
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• 제28권9호
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• pp.1499-1509
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• 2007

Proteomic analyses of synaptic vesicle fraction from rat brain have been performed for the better understanding of vesicle regulation and signal transmission. Two different approaches were applied to identify proteins in synaptic vesicle fraction. First, the isolated synaptic vesicle proteins were treated with trypsin, and the resulting peptides were analyzed using a high-pressure capillary reversed phase liquid chromatography/tandem mass spectrometry (cRPLC/MS/MS). Alternatively, proteins were separated by two-dimensional gel electrophoresis (2DE) and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF/MS). Total 18 and 52 proteins were identified from cRPLC/MS-MS and 2DE-MALDI-TOF-MS analysis, respectively. Among them only 2 proteins were identified by both methods. Of the proteins identified, 70% were soluble proteins and 30% were membrane proteins. They were categorized by their functions in vesicle trafficking and biogenesis, energy metabolism, signal transduction, transport and unknown functions. Among them, 27 proteins were not previously reported as synaptic proteins. The cellular functions of unknown proteins were estimated from the analysis of domain structure, expression profile and predicted interaction partners.

• The Korean Journal of Physiology
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• 제22권2호
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• pp.281-293
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• 1988

가토 신장 근위세뇨관에서 분리한 brush border membrane vesicle(BBMV)과 basolateral membrane vesicle(BBMV)에서 rapid filtration 방법으로 PAH 이동에 대한 pH의 영향을 관찰하였다. BLMV에서 용액내 Na이 없을 때 외부 $pH(pH_0)$를 8.0에서 5.5까지 감소시켰을 때 probenecid-sensitive PAH 이동은 유의하게 증가되었다. 용액내 Na이 있을 때 $pH_0$가 8.0에서 6.0까지 변화하여도 PAH 이동에는 영향이 없었으나 5.5까지 더욱 감소시켰을 때 PAH 이동이 증가하였다. 그러나 vesicle 내 외에 pH gradient없이 $pH_0$를 내부 $pH(pH_1)$와 동일하게 변화시켰을 때 PAH 이동은 영향을 받지 않았다. PH gradient가 없을 때 시험된 pH범위에서 Na은 PAH 이동을 증가시켰다. BBMV에서도 BLMV에서와 유사한 pH 의존성을 보였으나 Na의 존재는 PAH 이동에 영향을 미치지 못하였다. BLMV에서 동력학적 분석 결과 일정한 $pH(pH_1)$에서 $pH(pH_0)$ 감소는 Km에 변화없이 PAH 이동에 대한 Vmax를 유의하게 증가시켰다. 이러한 결과로 BBMV에서 PAH 이동에 대한 pH의 영향은$OH^-/PAH$교환기전에 기인하는 것으로 추측되나 BLMV에서 pH 의존성은 음이온 교환기전만으로 설명될 수 없다. 또한 BLMV에서는 PAH 이동이 Na에 의존하나 BBMV에서는 Na에 의존하지 않음을 가르킨다.

• The Korean Journal of Physiology
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• 제21권2호
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• pp.273-282
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• 1987

가토 신피질에서 분리한 brush border membrane (BBM)과 basolateral membrane vesicle (BLM)에서 유기 음이온인 p-aminohippuric acid (PAH)와 유기 양이온인 tetraethylammonium (TEA)의 이동에 대한 동력학적 분석을 하였다. BLM에서 PAH에 대한 Km과 Vmax값은 각각 $0.34{\pm}0.02\;mM$과 $0.22{\pm}0.07\;nmol/mg\;protein/20s$였으며 BBM에서 각 값은 $8.46{\pm}0.57\;mM$과 $4.43{\pm}0.40\;nmol/mg\;protein/20s$였다. BLM에서 용액내 Na의 제거는 PAH에 대한 Km 값에는 영향없이 Vmax 값을 변화시켰다. BBM에서 TEA이동에 대한 Km과 Vmax 값은 각각 $0.55{\pm}0.15\;mM$과 $1.04{\pm}0.23\;nmol/mg\;protein/20s$였으며 BLM에서 각 값은 $0.46{\pm}0.04\;mM$과 $0.61{\pm}0.06\;nmol/mg\;protein/20s$였다. BLM에서 측정한 유기 이온들의 이동에 대한 Km 값이 신절편이나 분리된 tubule에서 보고된 값과 일치함을 보였으며 이러한 결과는 신세뇨관 세포막을 통한 유기 이온들의 이동 특성이 membrane vesicle을 분리하는 과정에서 변하지 않았음을 가르킨다.

• Journal of Life Science
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• 제12권2호
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• pp.43-46
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• 2002

Kinesin Superfamily Protein 1A (KIF1A) is an anterograde monomeric motor transporting a subset of synaptic vesicle precursors and plays an important role in neuronal function and survival. Here, f have used the yeast two-hybrid system to identify the proteins that interacts with the tail region of KIF1A. Calmodulin was found to interact specifically with the tail region of KIF1A. Calmodulin regulates many diverse cellular functions by modulating the activity of the proteins that interact with it. KIF1A interacts with calmodulin in the yeast two-hybrid assay, which is proved by immunoprecipitation with calmodulin in brain fraction. These results indicate that KIF1A is associated with calmodulin, suggesting that calmodulin may be a key role in the regulation of anterograde transport of synaptic 1 vesicle precursors.

• 대한약리학회지
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• 제22권2호
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• pp.151-156
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• 1986

최근 심근세포 또는 신경세포에서 발표된 여러 종류의 calcium channel중 calcium antagonist로 차단되는 channel 또는 차단되지 않는 channel 등이 있는지 알아보기 위해 실험을 시행하였다. 돼지 소장 평활근으로부터 고농도의 KCl(150mM)로 부하된 세포 포막낭을 만들어 고농도의$K^+$ 또는 전기자극으로 $^{45}Ca$의 이동을 유발시켜 다음과 같은 성적을 얻었다. 저농도의 $K^+$용액에서의 $^{45}Ca$이동보다 고농도의 $K^+$-용액에서의 $^{45}Ca$이동이 유의하게 증가되었으며(p<0. 05) 이때 유입되는 $^{45}Ca$의 양은 시간에 따라 서서히 감소되었다. 전기자극(3V, 15Hz, 25msec)을 하였을때 유입되는 $^{45}Ca$의 양은 전기자극을 하지 않은 대조군에 비하여 현저하게 증가되었고, 자극시간에 따른 $^{45}Ca$의 유입량은 2분 동안 계속 증가되었다. Diltiazem 또는 nifedipine을 처치하였을때, 고농도의 $K^+$-용액에 의한 $^{45}Ca$의 유입은 억제되지 않았으나 전기자극에 의해 유도되는 $^{45}Ca$의 유입은 유의하게 억제되었다(p<0.005). 상기의 실험성적으로 돼지 소장 평활근으로부터 분리한 세포막에서의 calcium이동 중 전기자극에 의해 이루어지는 것은 calcium antagonist로 차단되는 calcium channel을 통하여 이루어지는 것으로 사료된다.

• Journal of Pharmaceutical Investigation
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• 제37권5호
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• pp.311-314
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• 2007

Paclitaxel (PTX) is an antineoplastic agent approved for the treatment of ovarian and breast carcinomas. However, the use of paclitaxel as an anticancer drug is limited by its extremely poor water solubility (below $0.3\;{\mu}g/mL$). Furthermore, it has very low bioavailability when administered orally because paclitaxel is a substrate of P-glycoprotein (P-gp) efflux pump. In this study, buccal delivery of PTX was investigated as one of the alternatives for PTX delivery. Ethanol and glyceryl monooleate (GMO) were selected as permeation enhancing agents to increase solubility and permeation across buccal mucosa of PTX. At the different concentrations of ethanol solution ($30{\sim}70\;w/w\;%$), PTX permeation was studied, followed effects of GMO in the concentration range of $2.5{\sim}25%$ with ethanol vesicle. The transbuccal ability of PTX was evaluated in vitro using Franz diffusion cells mounted with rabbit buccal mucosa. As a result, incorporation of PTX into ethanol vesicle with GMO significantly enhanced the PTX permeation in rabbit buccal mucosa. Particularly, the mixtures of ethanol:water:GMO at the ratio of 50:47.5:2.5 showed the most excellent enhancing ability. The results showed a promising possibility for buccal delivery of PTX.