• Title/Summary/Keyword: transmembrane

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Chromophore formation and phosphorylation analysis of constitutively active rhodopsin mutants (Chromophore 형성과 rhodopsin kinase 활성을 이용한 항활성 로돕신 mutant의 분석)

  • Kim, Jong-Myoung
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.783-790
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    • 2007
  • G protein coupled receptors (GPCRs) transmit various extracellular signals into the cells. Upon binding of the ligands, conformational changes in the extracellular and/or transmembrane (TM) domains of CPCRs were propagated into the cytoplasmic (CP) domain of the molecule leading to the activation of their cognate heterotrimeric C proteins and kinases. Constitutively active GPCR mutants causing the activation of C Protein signaling even in the absence of ligand binding are of interest for the study of activation mechanism of GPCRs. Two classes of constitutively active mutations, categorized by their effects on the salt bridge between Ell3 and K296, were found in the TM domain of rhodopsin. Opsin mutants containing combinations of the mutations were constructed to study the conformational changes required for the activation of rhodopsin. Rhodopsin chromophore regenerated with 11-cis-retinal showed a thermal stability inversely correlated with its constitutive activity. In contrast, rhodopsin mutants exhibited a binding affinity to an agonist, all-trans-retinal, in a constitutive activity-dependent manner. In order to test whether the conformational changes responsible for the activation of trans-ducin (Gt) are the same as the conformation required for the recognition of rhodopsin kinase, analysis of the mutants were carried out with phosphorylation by rhodopsin kinase. Rhodopsin mutants containing combinations of different classes of the mutations showed a strong synergistic effect on the phosphorylation of the mutants in the dark as similar to that of Gt activation. The results suggest that at least two or three kinds of segmental and independent conformational changes are required for the activation of rhodopsin and the conformational changes responsible for activating rhodopsin kinase and Gt are similar to each other.

Identification of cis-Regulatory Region Controlling Semaphorin-1a Expression in the Drosophila Embryonic Nervous System

  • Hong, Young Gi;Kang, Bongsu;Lee, Seongsoo;Lee, Youngseok;Ju, Bong-Gun;Jeong, Sangyun
    • Molecules and Cells
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    • v.43 no.3
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    • pp.228-235
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    • 2020
  • The Drosophila transmembrane semaphorin Sema-1a mediates forward and reverse signaling that plays an essential role in motor and central nervous system (CNS) axon pathfinding during embryonic neural development. Previous immunohistochemical analysis revealed that Sema-1a is expressed on most commissural and longitudinal axons in the CNS and five motor nerve branches in the peripheral nervous system (PNS). However, Sema-1a-mediated axon guidance function contributes significantly to both intersegmental nerve b (ISNb) and segmental nerve a (SNa), and slightly to ISNd and SNc, but not to ISN motor axon pathfinding. Here, we uncover three cis-regulatory elements (CREs), R34A03, R32H10, and R33F06, that robustly drove reporter expression in a large subset of neurons in the CNS. In the transgenic lines R34A03 and R32H10 reporter expression was consistently observed on both ISNb and SNa nerve branches, whereas in the line R33F06 reporter expression was irregularly detected on ISNb or SNa nerve branches in small subsets of abdominal hemisegments. Through complementation test with a Sema-1a loss-of-function allele, we found that neuronal expression of Sema-1a driven by each of R34A03 and R32H10 restores robustly the CNS and PNS motor axon guidance defects observed in Sema-1a homozygous mutants. However, when wild-type Sema-1a is expressed by R33F06 in Sema-1a mutants, the Sema-1a PNS axon guidance phenotypes are partially rescued while the Sema-1a CNS axon guidance defects are completely rescued. These results suggest that in a redundant manner, the CREs, R34A03, R32H10, and R33F06 govern the Sema-1a expression required for the axon guidance function of Sema-1a during embryonic neural development.

The Effect of Turbulence Promoters on the Mass Transfer in Ultrafiltration (한외여과에서의 물질전달에 대한 난류촉진물체의 영향)

  • Oh, Won-Suhk;Park, Ham-Yong;Lim, Gio-Bin;Kim, Woo-Sik
    • Membrane Journal
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    • v.4 no.4
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    • pp.221-231
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    • 1994
  • The GR51PP(MWCO 50,000) and GR40PP(MWCO 100,000) membranes manufactured by DDS were used in ultrafiltration of dextran(Mw. : 500,000) solution in flat plate ultrafiltration cell filled with various types of turbulence promoters. The flux improvement by using turbulence promoter was higher in laminar flow region than in turbulent flow region. The maximum improvements of permeate flux were foud as 112% and 50% I laminar flow region and turbulent flow region, respectively. Also, the solute rejection of the ultrafiltration membrane was improved by turbulence promoters and its effect was significant in the high transmembrane pressure and laminar flow region. The smaller the spacer mesh size was used, the higher the flux improved, but the pressure drop in ultrafiltration cell also increased. In laminar flow region, pressure drop by the spacer was negligible, but in turbulent flow region it changed significantly depending upon the mesh size of the spacer and therefore, its mesh size must be baken into account in the design of the process. The predicted results of the modified mass transfer correlation had better agreement with experimental results than those of unmodified one, The modified mass transfer correlations for laminar and turbulent flow region are shown as follow. $N_{sh}=0.151(N_{Re})^{0.199}(N_{Sc})^{0.22}(N_{Scm})^{0.197}\;(625 $N_{sh}=0.0165(N_{Re})^{0.428}(N_{Sc})^{0.33}(N_{Scm})^{0.223}\;(5015

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Effect of Cross-flow Velocity and TMP on Membrane Fouling in Thermophilic Anaerobic Membrane Bioreactor Treating Food Waste Leachate (음식물 침출수를 처리하는 막결합 고온혐기성 소화시스템에서 교차여과와 막간압력이 파울링에 미치는 영향)

  • Kim, Young-O;Jun, Duk-Woo;Yoon, Seong-Kyu;Chang, Chung-Hee;Bae, Jae-Ho;Yoo, Kwan-Sun;Kim, Jeong-Hwan
    • Membrane Journal
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    • v.21 no.4
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    • pp.360-366
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    • 2011
  • The effect of cross-flow velocity and transmembrane pressure (TMP) on membrane fouling was observed from pilot-scale operation of thermophilic anaerobic membrane bioreactor (AnMBR) treating food waste leachate. It was found that fouling rate was reduced significantly as cross-flow velocity increased at constant TMP mode of operation while this effectiveness was more pronounced at lower TMP. Higher TMP resulted in less permeable fouling layer possibly due to compressibility of foulant material on membrane surface. Particle sizes of membrane concentrate ranged from 10 to $100{\mu}m$, implying that shear-induced diffusion enhance back transport of these particle sizes away from the membrane effectively. From the continuous operation of AnMBR, it was confirmed that shear rate played an important role in the reduction of membrane fouling. Membrane autopsy works at the end of operation of AnMBR showed clearly that both organic and inorganic fouling were significant on membrane surface. Surface shear by cross-flow velocity was expected to be less effective to remove irreversible fouling which can be mainly caused by the adsorption of organic colloidal materials into membrane pores.

Concentration of Functional Mineral by NF/RO Processes (나노여과/역삼투 공정을 이용한 기능성 미네랄의 농축)

  • Lee, Ho-Won;Moon, Soo-Hyoung;Ko, Kyoung-Soo
    • Membrane Journal
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    • v.19 no.4
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    • pp.277-284
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    • 2009
  • In order to select the most suitable membrane to the concentration of vanadium and silica in groundwater, two different commercial NF membrane modules (NE2540-90 and NF90-2540) and three different commercial RO membrane modules (BW30-2540, RE2540-TE, and XLE-2540) were tested. The membrane characteristics test results showed that NE2540-90 module was the most efficient because of higher permeate flux and similar rejection coefficient. Using NE2540-90 module at the transmembrane pressure of $8\;kgf/cm^2$, it was found that the rejection coefficients of vanadium, silica, aluminium, chromium, iron, boron, strontium, and barium were 98.2%, 99.0%, 92.0%, 83.6%, 96.0%, 45.1%, 98.6%, and 69.5%, respectively. It was possible that vanadium and silica contents of groundwater were concentrated into $148.9\;{\mu}g/L$ and 85.8 mg/L respectively by six-stages NF process at the recovery ratio of 15%. The waters produced by NF, which are enriched in vanadium and silica content, are expected to be commercialized the various functional mineral waters.

Synthesis of (4-$[^{18}F]$Fluorophenyl)triphenylphosphonium as a Mitochondrial Voltage Sensor for PET (PET영상용 미토콘드리아 막전위 감지기 (4-$[^{18}F]$Fluorophenyl)triphenylphosphonium의 합성)

  • Kim, Dong-Yeon;Yu, Kook-Hyun;Bom, Hee-Seung;Min, Jung-Joon
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.6
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    • pp.561-565
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    • 2007
  • Purpose: Lipophilic cations including tetraphenylphosphonium (TPP) salts penetrate the hydrophobic barriers of the plasma and mitochondrial membranes, resulting in accumulation in mitochondria in response to the negative inner transmembrane potentials. The development of radiolabeled phosphonium cations as a noninvasive imaging agent may serve as a new molecular "voltage sensor" probe to investigate the role of mitochondria in the pathophysiology and diagnosis of cancer. Materials and Methods: We have synthesized a reference compound (4-fluorophenyl)triphenylphosphonium (TPP) and a labeled compound $[^{18}F]$TPP via two step nucleophilic substitution of no-carrier-added $[^{18}F]$fluoride with the precursor, 4-iodophenyltrimethylammonium iodide, in the presence of Kryptofix-2.2.2 and $K_2CO_3$. Result: The reference compound (4-fluorophenyl)triphenylphosphonium (TPP) was synthesized in 60% yield. The radiolabeled compound $[^{18}F]$TPP was synthesized in $10\sim15%$ yield. The radiochemical purity of the $[^{18}F]$TPP was $95.57{\pm}0.51%$ (n=11). Conclusion: $[^{18}F]$TPP was successfully synthesized that might have a potential to be utilized as a novel myocardial or cancer imaging agent for PET. However, it is required to improve the radiochemical yield to apply $[^{18}F]$TPP in preclinical or clinical researches.

Effect of TSHAC on Human Cytochrome P450 Activity, and Transport Mediated by P-Glycoprotein

  • Im, Yelim;Kim, Yang-Weon;Song, Im-Sook;Joo, Jeongmin;Shin, Jung-Hoon;Wu, Zhexue;Lee, Hye Suk;Park, Ki Hun;Liu, Kwang-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1659-1664
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    • 2012
  • TSAHC [4'-(p-toluenesulfonylamido)-4-hydroxychalcone] is a promising antitumorigenic chalcone compound, especially against TM4SF5 (four-transmembrane L6 family member 5)-mediated hepatocarcinoma. We evaluated the potential of TSAHC to inhibit the catalytic activities of nine cytochrome P450 isoforms and of P-glycoprotein (P-gp). The abilities of TSAHC to inhibit phenacetin O-deethylation (CYP1A2), coumarin 6-hydroxylation (CYP2A6), bupropion hydroxylation (CYP2B6), amodiaquine N-deethylation (CYP2C8), diclofenac 4-hydroxylation (CYP2C9), omeprazole 5-hydroxylation (CYP2C19), dextromethorphan O-demethylation (CYP2D6), chlorzoxazone 6-hydroxylation (CYP2E1), and midazolam 1'-hydroxylation (CYP3A) were tested using human liver microsomes. The P-gp inhibitory effect of TSAHC was assessed by [$^3H$]digoxin accumulation in the LLCPK1-MDR1 cell system. TSAHC strongly inhibited CYP2C8, CYP2C9, and CYP2C19 isoform activities with $K_i$ values of 0.81, 0.076, and $3.45{\mu}M$, respectively. It also enhanced digoxin accumulation in a dose-dependent manner in the LLCPK1-MDR1 cells. These findings indicate that TSAHC has the potential to inhibit CYP2C isoforms and P-gp activities in vitro. TSAHC might be used as a nonspecific inhibitor of CYP2C isoforms based on its negligible inhibitory effect on other P450 isoforms such as CYP1A2, CYP2A6, CYP2B6, CYP2D6, CYP2E1, and CYP3A.

Fouling Mitigation for Pressurized Membrane of Side-Stream MBR Process at Abnormal Operation Condition (가압식 분리막을 이용한 Side-Stream MBR 공정의 비정상 운전조건에서 막 오염 저감)

  • Ko, Byeong-Gon;Na, Ji-Hun;Nam, Duck-Hyun;Kang, Ki-Hoon;Lee, Chae-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.38 no.6
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    • pp.323-328
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    • 2016
  • Pressurized membrane used for side-stream MBR process requires fouling control strategy both for normal and abnormal operation conditions for stable operation of the facilities. In this study, $85m^3/day$ of pilot-scale side-stream MBR process was constructed for the evaluation of fouling mitigation by air bubble injection into the membrane module. In addition, fouling phenomena at abnormal operation conditions of low influent and/or loading rate were also investigated. Injection of air bubble was found to be effective in delaying transmembrane pressure (TMP) increase mainly due to scouring effect on the membrane surface, resulting in expanded filtration cycle at a high flux of $40L/m^2{\cdot}h$ (LMH). At abnormal operation condition, injection of PACl (53 mg/L as Al) into the bioreactor showed 19% reduction of TMP increase. However, inhibition of nitrifying bacteria by continuous PACl injection was observed from batch experiments. In contrast, injection of powdered activated carbon (PAC, 0.6 g/L) was able to maintain the initial TMP of $0.2kg/cm^2$ for 5 days at the abnormal conditions. It may have been caused from the adsorption of extracellular polymeric substances (EPS), which was known to be excessively released during growth inhibition condition and act as the major foulants in MBR operations.

Membrane Fouling Control Effect of Periodic Water-back-flushing in the Tubular Carbon Ceramic Ultrafiltration System for Recycling Paper Wastewater (제지폐수 재활용을 위한 관형 탄소계 세라믹 한외여과장치에서 물 역세척의 막오염 제어 효과)

  • 김미희;박진용
    • Membrane Journal
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    • v.11 no.4
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    • pp.190-203
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    • 2001
  • In this study the discharged wastewater from a paper plant was filtrated by 4 kinds of tubular carbon ceramic ultrafiltration membranes with periodic water-back-flushing. We could investigate effects of watch-back-flushing period, transmembrane pressure (TMP) and flow rate, and find optimal operating conditions. The back-f1ushing time (BT) was fixed at 3 sec, and fi1tration times (FT) werc changed in 15~60 scc, TMP in 1.00~2.50$kg_{f}$/$cm^2$, and the flow rates in 0.27~1.75 L/min. The optimal conditions were discussed in 7he viewpoints of dimensionless permeate flux (J/J$_{0}$), total permeate volume ($V^T$) and resistance of membrane fouling ($R^f$). Optima1 back-flushing period was BT/FT=0.20, suggesting that the frequent back-flushing should decrease membrane fouling. Optimal TMP in the viewpoint of $V^T$ was 1.00~1.55$kg_{f}$/$cm^2$, suggesting that rising TMP should increase membrane fouling and decrease permeate flux. But, rising f1ow rate should decrease membrane fouling and increase permeate flux. Then, average rejection rates of pollutants filtratedby carbon ceramic membranes were 88~98 % for turbidity, 48~72% fort $COD_{cr}$ and 37~76% for TDS.

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Effect of Periodic $N_2$-back-flushing in Paper wastewater Treatment using Carbon Ceramic Ultrafiltration and Microfiltration Membranes (탄소계 세라믹 한외 및 정밀 여과막으로 제지폐수 처리시 주기적 질소 역세척의 효과)

  • 황현정;박진용
    • Membrane Journal
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    • v.12 no.1
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    • pp.8-20
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    • 2002
  • In this study using $N_2$-back flushing, which wwas not the general back-flushing method of membranes, the discharged wastewater from a paper plant was filtrated by 4 kinds of tubular carbon ceramic ultrafiltration membranes. We could in vestigate effects of $N_2$-back flushing period, transmembrane pressure (TMP)and flow rate and find optimal operating conditions. The $N_2$-back flushing time (BT) was fixed at 40 sec, filtration times (FT) were changed in 4~32 min, TNP in $1.0~3.0kg_f/cm^2$ the flow celocities in 0.53~1.09cm/s. The optimal conditions were discussed in the viewpoints of dimensionless permeate flux ($J/J_0$), toal permeate volume ($V_T$) and resistance of membrane fouling ($R_f$). Optimal back-flushing period was BT/FT=0.167 (FT=8 min ), in which more $V_T$ was obtained than that in BT/FT=0.083 (FT=4 min) which was the most friquent back-flushing condition. Then rising TMP should increase the driving force, and more $V_T$ could be accumulated. And rising flow rate should decrease membrane fouling increase permeate flux, and more $V_T$could be produced. Average rejection rates of pollutants were higher than 95% for turbidity and 45~83% for $COD_{Cr}$, but rejection rates of total dissolved solid (TDS) were lower than 10%.