• Title/Summary/Keyword: Transport Pathway

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Genome-wide Analysis and Control of Microbial Hosts for a High-level Production of Therapeutic Proteins

  • Kim, Sung-Geun;Park, Jung-Hwan;Lee, Tae-Hee;Kim, Myung-Dong;Seo, Jin-Ho;Lim, Hyung-Kwon
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2005.06a
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    • pp.230-232
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    • 2005
  • The formation of insoluble aggregation of the recombinant kringle fragment of human apolipoprotein(a), rhLK8, in endoplasmic reticulum was identified as the rate-limiting step in the rhLK8 secretion in Saccharomyces cerevisiae. To analyze the protein secretion pathway, some of yeast genes closely related to protein secretion was rationally selected and their oligomer DNA were arrayed on the chip. The expression profiling of these genes during the induction of rhLK8 in fermentor fed-batch cultures revealed that several foldases including pdi1 gene were up-regulated in the early induction phase, whereas protein transport-related genes were up-regulated in the late induction phase. The coexpression of pdi1 gene increased rhLK8-folding capacity. Hence, the secretion efficiency of rhLK8 in the strain overexpressing pdi1 gene increased by 2-fold comparing in its parental strain. The oligomer DNA chip arrayed with minimum number of the genes selected in this study could be generally applicable to the monitoring system for the heterologous protein secretion and expression in Saccharomyces cerevisiae. With the optimization of fed-batch culture conditions and the alteration of genetic background of host, we obtained extracellular rhLK8 at higher yields than with Pichia pastoris systems, which was a 25-fold increased secretion level of rhLK8 compared to the secretion level at the initiation of this study.

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Enhancing Effect of Extracts of Phellodendri Cortex on Glucose Uptake in Normal and Insulin-resistant 3T3-L1 Adipocytes (3T3-L1 지방세포에서 황백 추출물의 Glucose Uptake 촉진 및 인슐린 저항성 개선 활성)

  • Kim, So-Hui;Shin, Eun-Jung;Hyun, Chang-Kee
    • Korean Journal of Pharmacognosy
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    • v.36 no.4 s.143
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    • pp.291-298
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    • 2005
  • Anti-hyperglycemic effects of 17 medicinal plants that have been used for ameliorating diabetes in oriental medicine were evaluated using glucose transport assay in 3T3-L1 adipocytes. Higher activities were obtained by treating water or alcohol extract of Phellodendri Cortex (PC), which showed enhancing effects both on basal and insulin-stimulated glucose uptake. The latter effect of PC was completely inhibited by wortmannin, a specific inhibitor for phosphatidyl inositol 3-kinase (PI 3-kinase), but not affected by SB203580, A specific inhibitor for p38 mitogen-activatedprotein kinase(MAPK). Genistein, an inhibitor for tyrosine kinases, abolished the PC effects completely. Treatment of vanadate, an inhibitor for tyrosine phosphatases, together with PC showed no significant synergic enhancement in glucose uptake. The results of inhibitors associated with insulin signaling pathway indicated that extracts of PC enhance glucose uptake by PI-3 kinase activation which is an upstream event for GLUT4 translocation. Antidiabetic effects of PC extract might be also due to enhanced tyrosine phosphorylation and reduced tyrosine dephosphorylation. In addition, PC accelerated insulin-stimulated glucose uptake in insulin-resistant cells, recovering the uptake level close to that of normal cells. These findings may offer a new way to utilize extracts of PC as novel anti-hyperglycemic agents.

RNAseq-based Transcriptome Analysis of Burkholderia glumae Quorum Sensing

  • Kim, Sunyoung;Park, Jungwook;Kim, Ji Hyeon;Lee, Jongyun;Bang, Bongjun;Hwang, Ingyu;Seo, Young-Su
    • The Plant Pathology Journal
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    • v.29 no.3
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    • pp.249-259
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    • 2013
  • Burkholderia glumae causes rice grain rot and sheath rot by producing toxoflavin, the expression of which is regulated by quorum sensing (QS). The QS systems of B. glumae rely on N-octanoyl homoserine lactone, synthesized by TofI and its cognate receptor TofR, to activate the genes for toxoflavin biosynthesis and an IclR-type transcriptional regulator gene, qsmR. To understand genome-wide transcriptional profiling of QS signaling, we employed RNAseq of the wild-type B. glumae BGR1 with QS-defective mutant, BGS2 (BGR1 tofI::${\Omega}$) and QS-dependent transcriptional regulator mutant, BGS9 (BGR1 qsmR::${\Omega}$). A comparison of gene expression profiling among the wild-type BGR1 and the two mutants before and after QS onset as well as gene ontology (GO) enrichment analysis from differential expressed genes (DEGs) revealed that genes involved in motility were highly enriched in TofI-dependent DEGs, whereas genes for transport and DNA polymerase were highly enriched in QsmR-dependent DEGs. Further, a combination of pathways with these DEGs and phenotype analysis of mutants pointed to a couple of metabolic processes, which are dependent on QS in B. glumae, that were directly or indirectly related with bacterial motility. The consistency of observed bacterial phenotypes with GOs or metabolic pathways in QS-regulated genes implied that integration RNAseq with GO enrichment or pathways would be useful to study bacterial physiology and phenotypes.

Physiology of Anthocyanin Accumulation in Corn Leaves Treated with Metsulfuron-methyl (Metsulfuron-methyl 처리에 의한 옥수수의 Anthocyanin 축적 생리에 관한 연구)

  • Park, E.Y.;Kim, J.S.;Cho, K.Y.;Pyon, J.Y.
    • Korean Journal of Weed Science
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    • v.18 no.3
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    • pp.246-256
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    • 1998
  • The objective of this study was to investigate the cause of anthocyanin accumulation in the corn leaves treated with metsulfuron-methyl. The accumulation of anthocyanin and total sugar was increased with the time after metsulfuron-methyl application and with the greater herbicide concentration as well. The anthocyanin increase was alleviated by the combined treatment of DUMU or the tank-mixture treatment of metsulfuron-methyl, isoleucine and valine. Metsulfuron-methyl could not enhance the anthocyanin formation in the nonchlorophyllous tissue in which photosynthetic carbohydrate production was limited. Upon the exogenous supply of sucrose, however, the contents of anthocyanin was increased in the both chlorophyllous and nonchlorophyllous leaf segment of corn without the herbicide treatment. On the other hand, the herbicide itself did not influence Hill reaction in vitro and photosynthesis electron transport in vivo. The anthocyanin accumulation by metsulfitronmethyl did not occur in the corn mutants deficient in the structural gene of anthocyanin synthesis, but corns deficient in the regulatory gene had the anthocyanin accumulation only in R-r and r-r type. The above results suggest that the purple pigmentation in the corn leaves treated with metsulfuron-methyl is related to the accumulation of photosynthetic carbohydrate which can stimulate the. regulatory gene related to anthocyanin biosynthetic pathway.

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Understanding of Growth Habits of $VO_2$ Film on Graphene and Their Effects on Metal to Insulator $Transition_2$

  • Yang, Jae-Hoon;Kim, Keun-Soo;Jang, A-Rang;Yang, Hyoung-Woo;Kang, Dae-Joon
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.572-572
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    • 2012
  • Growth of metal oxides on graphene may lead to a better understanding of delicate effects of their growth habits on their underlying physics. The vanadium dioxide ($VO_2$) is well known for its metal-to-insulator transition accompanied by a reversible first order structural phase transition at 340 K. This transition makes $VO_2$ a potentially useful material for applications in electrical and optical devices. We report a successful growth of $VO_2$ nanostructures on a graphene substrate via a vapor-solid transport route. As-grown $VO_2$ nanostructures on graphene were systematically characterized by field emission scanning electron microscopy, x-ray diffraction, Raman spectroscopy, FT-IR spectroscopy and high resolution transmission electron microscopy. These results indicate that the strain between $VO_2$ and graphene layers may be easily controlled by the number of underlying graphene layer. We also found that the strain in-between $VO_2$ and graphene layer affected its metal-to-insulator transition characteristics. This study demonstrates a new way for synthesizing $VO_2$ in a desired phase on the transparent conducting graphene substrate and an easy pathway for controlling metal-to-insulator phase transition via strain.

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Assessment of Climate Change Impact on Best Management Practices of Highland Agricultural Watershed under RCP Scenarios using SWAT (SWAT모형을 이용한 RCP 기후변화 시나리오에 따른 고랭지농업유역의 최적관리기법 평가)

  • Jang, Sun-Sook;Kim, Seong-Joon
    • Journal of The Korean Society of Agricultural Engineers
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    • v.60 no.4
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    • pp.123-132
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    • 2018
  • The purpose of this study was to evaluate the reduction effect of non point source (NPS) pollution in Haean highland agricultural catchment ($62.8km^2$) for 13 BMP scenarios under RCP (Representative Concentration Pathway) 4.5 and 8.5 scenarios. Under the present climate condition, the BMP (best management practices) reduction efficiency of SS (suspended solid), T-N (total nitrogen), and T-P (total phosphorus) showed +25.7%, +4.2%, and +16.1% for VFS (vegetative filter strip), +0.1%, +15.6%, and +5.7% for FC (fertilizer control), and +6.3%, -2.9%, and +3.9% for RSM (rice straw mulching) respectively. In general, effective was the best for SS and T-P reductions, and the FC was the best for T-N reduction. The negative effect of T-N on RSM was induced by increase in infiltration and solute transport to baseflow. Under the future climate change scenarios, the SS, T-N, and T-P reduction efficiency showed the range of +1.9~+11.6%, -1.9~+0.2%, and +5.3~+11.9% respectively. The 3 BMPs (VFS, FC, and RSM) application in the future showed negative and little differences (-0.5~+1.6%) for SS and T-N reduction efficiencies while T-P reduction efficiency showed +0.3~+7.6% comparing with the baseline period. To achieve an increase in the reduction efficiency of future SS and T-N by +2~+10%, the combined application of more than two BMPs is necessary.

Regulations of Reversal of Senescence by PKC Isozymes in Response to 12-O-Tetradecanoylphorbol-13-Acetate via Nuclear Translocation of pErk1/2

  • Lee, Yun Yeong;Ryu, Min Sook;Kim, Hong Seok;Suganuma, Masami;Song, Kye Yong;Lim, In Kyoung
    • Molecules and Cells
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    • v.39 no.3
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    • pp.266-279
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    • 2016
  • The mechanism by which 12-O-tetradecanoylphorbol-13-acetate (TPA) bypasses cellular senescence was investigated using human diploid fibroblast (HDF) cell replicative senescence as a model. Upon TPA treatment, protein kinase C (PKC) ${\alpha}$ and $PKC{\beta}1$ exerted differential effects on the nuclear translocation of cytoplasmic pErk1/2, a protein which maintains senescence. $PKC{\alpha}$ accompanied pErk1/2 to the nucleus after freeing it from $PEA-15pS^{104}$ via $PKC{\beta}1$ and then was rapidly ubiquitinated and degraded within the nucleus. Mitogen-activated protein kinase docking motif and kinase activity of $PKC{\alpha}$ were both required for pErk1/2 transport to the nucleus. Repetitive exposure of mouse skin to TPA downregulated $PKC{\alpha}$ expression and increased epidermal and hair follicle cell proliferation. Thus, $PKC{\alpha}$ downregulation is accompanied by in vivo cell proliferation, as evidenced in 7, 12-dimethylbenz(a)anthracene (DMBA)-TPA-mediated carcinogenesis. The ability of TPA to reverse senescence was further demonstrated in old HDF cells using RNA-sequencing analyses in which TPA-induced nuclear $PKC{\alpha}$ degradation freed nuclear pErk1/2 to induce cell proliferation and facilitated the recovery of mitochondrial energy metabolism. Our data indicate that TPA-induced senescence reversal and carcinogenesis promotion share the same molecular pathway. Loss of $PKC{\alpha}$ expression following TPA treatment reduces pErk1/2-activated SP1 biding to the $p21^{WAF1}$ gene promoter, thus preventing senescence onset and overcoming G1/S cell cycle arrest in senescent cells.

Characterization of Methanol Crossover through Nafion Membranes by Direct Cell Performance Measurement

  • Park, Kyung-Won;Kim, Young-Min;Kwon, Bu-kil;Choi, Jong-Ho;Park, In-Su;Sung, Yung-Eun
    • Journal of the Korean Electrochemical Society
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    • v.5 no.4
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    • pp.226-231
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    • 2002
  • Power densities produced by the permeation of methanol through membranes were directly measured by inserting the membrane in front of anode in a membrane-electrode-assembly of a direct methanol fuel cell (DMFC). The power density was closely related to the loss of power in the DMFC and was strongly affected by temperature. As the cell temperature was increased, the power density resulting from methanol crossover was increased. The increase in methanol crossover had be attributed to diffusion caused or affected by temperature. Methanol crossover a major effect on the performance of a DMFC at a relatively low temperature with $26\%\;loss\;at\;30^{\circ}C$. In order to reduce methanol crossover, a conventional Nafion membrane was modified by the incorporation of Pt or Pd. The reduction in methanol crossover was investigated in these modified membranes by our cell performance measurement. Pt and Pd particles incorporated in the Nafion membranes block methanol pathway and prevent methanol transport through the membranes, which was proved by combining with liquid chromatography.

Glucose Transporters and AMP-Activated Protein Kinase Modulation Effects of Decursin and Decursinol Angelate on Diabetic Rats (당뇨유발 흰쥐에서 당수송 인자와 AMP-Activated Protein Kinase의 조절에 대한 데커신과 데커시놀 안젤레이트의 효과)

  • Ok, Seon;Lee, Ju-Hee;Kim, Ik-Hwan;Kang, Jae-Seon
    • YAKHAK HOEJI
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    • v.55 no.4
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    • pp.301-308
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    • 2011
  • Diabetes has been one of major health risks in industrialized countries. AMP-activated protein kinase (AMPK) has been focused as a novel therapeutic target for the treatment of metabolic syndromes, because AMPK increases glucose uptake through independent insulin signal pathway. In this study, we investigated the anti-diabetic effect of Angelica gigas Nakai extract (AGNEX), a mixture of decursin and decursinol angelate (53 : 47), decursin and decursinol angelate on blood glucose, glucose transport (GLUT) and AMPK expression levels in streptozotocin (STZ)-induced diabetic rats. To induce diabetes, 50 mg/kg of STZ was injected via i.v. route and AGNEX 2 mg/kg (STZ+AG), decursin 2 mg/kg (STZ+D), decursinol angelate 2 mg/kg (STZ+DA), and metformin 100 mg/kg (STZ+M) were administered orally for 21 days. STZ+DA group showed a significant decrease in fasting blood glucose levels compared to the other groups. Decursinol angelate significantly upregulated expression of glucose transporter 4 (GLUT4) and phosphorylation of AMPK (p-AMPK) in skeletal muscle of rats. In pancreas of rats, decursinol angelate significantly increased expression of GLUT2 through down-regulation of p-AMPK. In addition to the result of pancreatic islets morphology, AGNEX, decursin, decursinol angelate, and metformin treated group recovered ${\beta}$-cell damage by hyperglycemia. These results indicate that decursinol angelate might be a potential anti-diabetic agent and AGNEX could be useful in the treatment of diabetes mellitus.

Glu-56 in Htrl is Critical for Phototaxis Signaling in Halobacterium salinarum

  • Choi, Ah-Reum;Kim, So-Young;Yoon, Sa-Ryong;Jung, Kwang-Hwan
    • Animal cells and systems
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    • v.9 no.3
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    • pp.139-144
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    • 2005
  • The attractant (orange light) or repellent (white light) signal is transmitted from SRI (Sensory Rhodopsin I) via protein-protein interaction with its transducer Htrl (Halobacterial Transducer for Sensory Rhodopsin I) which in turn controls a cytoplasmic phospho-transfer pathway that modulates flagella motor switching in Halobacterium salinarum. Some mutations in both SRI and Htrl showed an unusual mutant phenotype called inverted signaling, in which the cell produces a repellent response to normally attractant light. Twelve mutations at the Glutamate 56 (E56) position in the second transmembrane helix of Htrl were introduced by site-specific random mutagenesis. Almost all E56 mutants showed orange-light inverted responses in pH and temperature-dependent manners except E56D and E56Y. Except for these two mutants, all mutants accelerated the $S_{373}$ decay compared to wild-type at $18^{\circ}C$. This supported that there is an interaction between SRI and the second transmembrane of Htrl. Also a structural model of Htrl based on the Tar crystal structure and the secondary structure prediction program proposed the E56 residue to be in the middle of the proton channel. The most important observation is that the E56 mutant provides the evidence that this residue is very sensitive for signal relay, which can be explained by the open and closed conformations of the channel (A and R conformations) in SRI, as was postulated by the unified conformational shuttling model for transport and signaling.