• Title/Summary/Keyword: spermidine transport

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Characterization of Spermidine Transport System in a Cyanobacterium, Synechocystis sp. PCC 6803

  • Raksajit, Wuttinun;Yodsang, Panutda;Maenpaa, Pirkko;Incharoensakdi, Aran
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.447-454
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    • 2009
  • The transport of spermidine into a cyanobacterium, Synechocystis sp. pec 6803, was characterized by measuring the uptake of $^{14}C$-spermidine. Spermidine transport was shown to be saturable with an apparent affinity constant ($K_m$) value of $67{\mu}M$ and a maximal velocity ($V_{max}$) value of 0.45 nmol/min/mg protein. Spermidine uptake was pH-dependent with the pH optimum being 8.0. The competition experiment showed strong inhibition of spermidine uptake by putrescine and spermine, whereas amino acids were hardly inhibitory. The inhibition kinetics of spermidine transport by putrescine and spermine was found to be noncompetitive with $K_i$ values of 292 and $432{\mu}M$, respectively. The inhibition of spermidine transport by various metabolic inhibitors and ionophores suggests that spermidine uptake is energy-dependent. The diminution of cell growth was observed in cells grown at a high concentration of NaCl. Addition of a low concentration of spermidine at 0.5 mM relieved growth inhibition by salt stress. Upshift of the external osmolality generated by either NaCl or sorbitol caused an increased spermidine transport with about 30-40% increase at 10 mosmol/kg upshift.

Effect of Polyamine on Auxin Polar Transport in Corn Coleoptile Segments (Polyamine이 옥수수 자엽초 절편에서 오옥신 이동에 미치는 영향)

  • 허윤정
    • Journal of Plant Biology
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    • v.34 no.4
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    • pp.297-302
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    • 1991
  • The effects of polyamines on auxin polar transport were studied in corn coleoptile segments. Among putresine, spermidine and spermine tested in labelled auxin transport, spermidine inhibited auxin polar transport most strongly. Its inhibitory effect appeared after 1 h of transport period. Spermidine inhibited labelled auxin and 14C-benzoic acid accumulation into the tissue in the various pH range tested (pH 4.0-8.0). These results suggest that the inhibition of auxin transport may not be due to decrease in pH by spermine the effect of decreased pH in the extracellular space.

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Analysis of Polyamine Transport of Young Spring Radish Cotyledons (무우의 유자엽에서 Polyamine의 수송 분석)

  • Cho, Bong-Heuy
    • Analytical Science and Technology
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    • v.11 no.2
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    • pp.130-134
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    • 1998
  • Polyamines were analyzed in young cotyledons of spring radish. The uptake rate of SPD were transported within 20 min lineary and reached the saturation phase after 1hr. The uptake rate of SPD decreased gradually with the time by the increasing amount of SPD inside the cells. The uptake of PA depends on the external pH. The optimal pH of PA uptake are pH 7.5. $K_m-$ and $V_{max}-$values depend on external pH also. The uptake rate of PA was inhibited by external KCI, which depolarized membrane potential in the cells.

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Putrescine Transport in a Cyanobacterium Synechocystis sp. PCC 6803

  • Raksajit, Wuttinun;Maenpaa, Pirkko;Incharoensakdi, Aran
    • BMB Reports
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    • v.39 no.4
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    • pp.394-399
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    • 2006
  • The transport of putrescine into a moderately salt tolerant cyanobacterium Synechocystis sp. PCC 6803 was characterized by measuring the uptake of radioactively-labeled putrescine. Putrescine transport showed saturation kinetics with an apparent $K_m$ of $92{\pm}10\;{\mu}M$ and $V_{max}$ of $0.33{\pm}0.05\;nmol/min/mg$ protein. The transport of putrescine was pH-dependent with highest activity at pH 7.0. Strong inhibition of putrescine transport was caused by spermine and spermidine whereas only slight inhibition was observed by the addition of various amino acids. These results suggest that the transport system in Synechocystis sp. PCC 6803 is highly specific for polyamines. Putrescine transport is energy-dependent as evidenced by the inhibition by various metabolic inhibitors and ionophores. Slow growth was observed in cells grown under salt stress. Addition of low concentration of putrescine could restore growth almost to the level observed in the absence of salt stress. Upshift of the external osmolality generated by either NaCl or sorbitol caused an increased putrescine transport with an optimum 2-fold increase at 20 mosmol/kg. The stimulation of putrescine transport mediated by osmotic upshift was abolished in chloramphenicol-treated cells, suggesting possible involvement of an inducible transport system.

Characterization of Putrescine Uptake in Hamster Amelanocytic Melanoma AMEL-3 Cells

  • Garcia-Fernandez, Antonio J.;Rodriguez, Rosa A.;Perez-Pertejo, Yolanda;Balana-Fouce, Rafael
    • Molecules and Cells
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    • v.20 no.1
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    • pp.127-135
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    • 2005
  • The uptake of putrescine, spermidine and spermine by Fortner's hamster amelanocytic melanoma AMEL-3 cells was observed in this study to be time-dependent, temperature-sensitive, pH-dependent and saturable. Metabolic poisons nullified polyamine uptake, an indication that this is an energy-requiring mechanism. The presence of $Na^+$ ions was found to be requisite to full activity. Valinomycin, gramicidin, monensin and the calcium ionophore calcimycin were also observed to inhibit the process substantially. The transporter active site would seem to contain sulfhydryl groups. Other diamines and polyamine analogues, as well as cationic diamidines, suppressed putrescine uptake. The presence of the ornithine decarboxylase inhibitor DFMO in the culture medium induced putrescine inflows. Putrescine, in turn, induced the negative expression of the carrier, thus suggesting that this influx mechanism is governed by up/down regulation. The cationic diamidine CGP 40215A and its analogue CGP039937A competitively inhibited putrescine transport, with Ki values of 1.9 and $15{\mu}M$, respectively. The role of polyamine uptake in these cultures is discussed.

Proteome Analysis of Paenibacillus polymyxa E681 Affected by Barley

  • Seul, Keyung-Jo;Park, Seung-Hwan;Ryu, Choong-Min;Lee, Yong-Hyun;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.934-944
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    • 2007
  • Paenibacillus polymyxa E681 is known to be able to suppress plant diseases by producing antimicrobial compounds and to promote plant growth by producing phytohormones, and secreting diverse degrading enzymes. In spite of these capabilities, little is known regarding the flow of information from the bacterial strain to the barley roots. In an attempt to determine the flow of information from the bacterial strain to barley roots, the strain was grown in the presence and absence of barley, and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MALDI-TOF mass spectrometry were used. 2D-PAGE detected approximately 1,000 spots in the cell and 1,100 spots in the supernatant at a pH 4-10 gradient. Interestingly, about 80 spots from each sample showed quantitative variations. Fifty-three spots from these were analyzed by MALDI-TOF mass spectrometry and 28 proteins were identified. Most of the cytosolic proteins expressed at higher levels were found in P. polymyxa E681 cells grown in the presence of barley rather than in the absence of barley. Proteins detected at a lower level in the surpernatant of P. polymyxa E68l cells grown in the presence of barley were lipoprotein, glucose-6-phosphate 1-dehydrogenase, heat-shock protein HtpG, spermidine synthase, OrfZ, ribonuclease PH, and coenzyme PQQ synthesis protein, and flagellar hook-associated protein 2 whereas proteins detected at a higher level in the surpernatant of P. polymyxa E681 cells grown in the presence of barley included D-alanyl-D-alanine ligase A, isopentenyl-diphosphate delta-isomerase, ABC transporter ATP-binding protein Uup, lipase. Many of the proteins belonging to plant-induced stimulons are associated with biosynthetic metabolism and metabolites of proteins and transport. Some of these proteins would be expected to be induced by environmental changes resulting from the accumulation of plant-secreted substances.

A Study on the Effective Container Transport System for the Relief of Urban Traffic Congestion -A Container Transport Time-Oriented- (도시교통 체증완화를 위한 효율적인 수송체계에 관한 연구 - 컨테이너 수송시간을 중심으로 -)

  • Kim, Tae-Gon
    • Journal of Korean Port Research
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    • v.4 no.1
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    • pp.33-43
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    • 1990
  • 우리나라에서는 70년대 이후 컨테이너 수송체계가 도입되어 운영되어 왔으나 차량의 급격한 증가와 더불어 컨테이너 수송체계는 도시교통에 있어서 많은 문제를 야기시키는 것으로 논란이 되고 있다. 특히, 부산항은 우리나라 최대의 국제 무역항으로써 전 컨테이너 수송화물의 90%이상을 처리하고 있고, 또한 수출입항으로서 제 역할을 다하고 있으나 낮은 도로율(12.45%)과 산재해 있는 30여개의 Off-Dock CY로 인해서 도심을 통과하는 컨테이너 수송차량은 많은 교통문제를 유발시키고 있다. 본 연구는 1) 부산시에 산재해 있는 30여개의 Off-Dock CY에 이르는 주요 도로상에서 컨테이너 수송량의 신간별 분포에 따라 컨테이너의 운행시간대를 첨두 시간대, 비첨두 시간대 및 심야 시간대로 분류하여 각 시간대별 평균 수송 시간 및 교통 체증으로 인한 수송 지체수준을 확인하였고, 2) 컨테이너 전용부두로 부터 Off-Dock CY에 이르는 도시 고속도로 및 주요간선도로상에서의 수송시간 및 수송 지체수준을 비교 분석하였으며, 3) 마지막으로, 효율적인 컨테이너 수송체계를 위한 최적 수송시간대 및 도로체계를 제시할 수 있었다. 특히, 컨테이너 수송시간 및 지체수준은 각 운행 시간대에 따라 현저한 차이를 보이고 있었는데, 심야 시간대를 이용하여 컨테이너 수송을 할 경우에는 다른 시간대에 비하여 50%정도의 수송시간 절감효과를 기대할 수 있었으며, 도시 고속도로를 이용할 경우에는 도시 간선도로에 비하여 30%정도의 수송시간을 절약할 수 있었다. 따라서 도시지역의 교통체중을 완화시키기 위해서 심야 시간대에 보다 많은 컨테이너 수송차량이 이용할 수 있는 컨테이너 수송체계가 확립되어야 하고, 산재해 있는 Off-Dock CY를 몇개의 ODCY그룹이나 단지까지 컨테이너 전용 고속도로의 건설이 바람직하고 생각된다.리구에서 조사한 결과 $10^{-3}$ M MGBG 단독 처리구에서 에틸렌 생성량이 가장 높았으며, spermidine과 spermine의 함량은 가장 낮았다. 부정근 형성능이 다소 회복된 $10^{-3}$ M MGBG+ $10^{-5}$M spermine 혼합처리구에서의 에틸렌 생성량은 대조구보다는 다소 높게 나타났다. 부정근 회복능이 가장 높았던 $10^{-3}$ M MGBG+ $10^{-5}$ M spermine + $10^{-4}$ M Cocl$_2$ 혼합처리구에서 에틸렌 생성량은 가장 낮게 나타났으며, polyamine 함량은 가장 높게 나타났다.(Cyperaceae), 명아주과(Chenopodiaceae) 같은 풀들이 하구언(estuary)의 해안가에 주로 서식하였던 것으로 나타났다. 이 시기동안에 해양성조류인 와편모류와 담수성 조류가 동시에 다양하게 산출되는 것은 하구언 퇴적환경이었음을 잘 지시해 준다. 제II화분대(고도, 43.72{\sim}3.36\;m$)가 시작되면서 혼합림은 침엽수림으로 바뀌었으며, 낙엽활엽수들의 서식지가 2엽소나무로 대치되면서 침엽수가 주요 수종을 이루었던 것으로 나타났다. 아마도 이런 산림형태는 자연 기후환경변화 뿐만 아니라 인간간섭에 의해 나타난 결과로 생각된다. 이 시기에 경작식물인 벼과(Gramineae), 옥수수(Zea), 메밀(Fagopyrum)과 논밭의 뚝 등지 에서 서식하는 돼지풀(Ambrosia) 등이 처음으로 산출되는 것으로 보아 인간간섭이 행해졌음을 잘 뒷 받침해준다. 또한 담수성 조류는 변함없이 산출되는 반면, 해양성조류인 와편모류가 산출되지 않는 것으로 보아 해안환경에서 다소 멀 어진

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