• 제목/요약/키워드: Binding state

검색결과 480건 처리시간 0.034초

Heat Shock Protein 90 Regulates the Stability of c-Jun in HEK293 Cells

  • Lu, Chen;Chen, Dan;Zhang, Zhengping;Fang, Fang;Wu, Yifan;Luo, Lan;Yin, Zhimin
    • Molecules and Cells
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    • 제24권2호
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    • pp.210-214
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    • 2007
  • The 90-kDa heat shock protein (HSP90) normally functions as a molecular chaperone participating in folding and stabilizing newly synthesized proteins, and refolding denatured proteins. The HSP90 inhibitor geldanamycin (GA) occupies the ATP/ADP binding pocket of HSP90 so inhibits its chaperone activity and causes subsequent degradation of HSP90 client proteins by proteasomes. Here we show that GA reduces the level of endogenous c-Jun in human embryonic kidney 293 (HEK293) cells in a time and dose dependent manner, and that this decrease can be reversed by transfection of HSP90 plasmids. Transfection of HSP90 plasmids in the absence of GA increases the level of endogenous c-Jun protein, but has no obvious affect on c-Jun mRNA levels. We also showed that HSP90 prolongs the half-life of c-Jun by stabilizing the protein; the proteasome inhibitor N-benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG132) blocks the degradation of c-Jun promoted by GA. Transfection of HSP90 plasmids did not obviously alter phosphorylation of c-Jun, and a Jun-2 luciferase activity assay indicated that over-expression of HSP90 elevated the total protein activity of c-Jun in HEK293 cells. All our evidence indicates that HSP90 stabilizes c-Jun protein, and so increases the total activity of c-Jun in HEK293 cells.

Self-Assembled ZnO Hexagonal Nano-Disks Grown by RF Sputtering

  • 정은지;김지현;김수진;강현철
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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    • pp.461-461
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    • 2013
  • Over the last decade, zinc oxide (ZnO) thin films have attracted considerable attention owing to large band gap of 3.37 eV and large exciton binding energy of 60 meV at room temperature [1-3]. Recent interest in ZnO related researches has been switched into the fabrication and characterization of low-dimensional nanostructures, such as nano-wires and nano-dots that can be applicable to manufacture the optoelectronic devices such as ultraviolet lasers, light-emitting-diodes and detectors. Since the optical properties of ZnO nano-structures might be distinct from those of bulk materials or thin films, the low-dimensional phenomena should be examined further. In order to utilize such advanced optoelectronic devices, one of the challenges is how to control the surface state related emissions that are drastically increased with increasing the density of the nano-structures and the surface-to-volume ratio. This paper reports the synthesis and characterization of self-assembled ZnO hexagonal nano-disks grown by radio-frequency magnetron sputtering. X-ray diffraction data and scanning electron microscopy data showed that ZnO hexagonal nano-disks were nucleated on top of the flat surfaces as the film thickness reached to 1.56 ${\mu}m$ and then the number of nano-disks increased with increasing the film thickness. The lateral size of hexagonal nano-disks was ~720 nm and height was ~74 nm. The strong photo luminescence spectra obtained at 10 K was also observed, which was assigned to a surface exciton emission at 3.3628 eV arising from the surface sites of hexagonal nano-disks.

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Gold-protein A Complex 항체 고정화법을 이용한 Salmonella spp.의 신속 검출 (Rapid Detection of Salmonella spp. by Antibody Immobilization with Gold-protein A Complex)

  • 박인선;김남수
    • 한국식품과학회지
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    • 제31권1호
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    • pp.1-6
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    • 1999
  • Salmonella spp.의 신속한 검출을 위하여 발진모듈, 수정결정 진동측정기, 박막형태의 수정결정으로 이루어진 압전류적(piezoelectric, PZ) 항체센서 시스템을 구성하였다. 수정결정의 금전극 표면에 Salmonella 구조 항원(common structural antigen)에 대한 항체를 protein A를 사용하여 고정화하고, 항체가 고정화된 수정결정과 미생물간의 결합반응에 의한 질량증가로 나타나는 진동수의 감소량을 측정하였다. PZ 항체센서는 $35^{\circ}C$, pH 7.2의 0.1M 인산 완충용액에서 Salmonella균에 대하여 가장 높은 감응도를 나타내었다. PZ 항체센서의 반응은 Salmonella균에 대하여 매우 선택적이었고 polystyrene bead 첨가시 센서의 감응도가 크게 증가하였다. Salmonella균의 농도가 $10^5{\sim}10^6\;CFU/mL$의 범위 내에 있을 때 쌍대수좌표에서 직선구간의 검량선을 얻을 수 있었고, Salmonella 검출에 소요되는 시간은 50분이내 이었다.

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Elucidation of Serpin's Conformational Switch Mechanism By Rapid Kinetic Study

  • Kang, Un-Beom;Lee, Cheolju;Baek, Je-Hyun;Seunghyun Ryu;Kim, Joon;Yu, Myeong-Hee
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.62-62
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    • 2003
  • The native form of serpin (serine protease inhibitor) is kinetically trapped in metastable state. Metastability in these proteins is critical to their biological function. Serpins inhibit target proteases by forming a stable covalent complex in which the cleaved reactive site loop of the serpin is inserted into $\beta$-sheet A of the serpin with concomitant translocation of the protease to the opposite of the initial binding site. Despite recent determination of the crystal structures of a Michaelis protease-serpin complex as well as a stable covalent complex, details on the kinetic mechanism remain unsolved. In this study we constructed several $\alpha$$_1$-antitrypsin variants and examined their kinetic mechanism of loop translocation and formation of protease-serpin complex by stopped-flow experiments of fluorescence resonance energy transfer as well as quenched-flow experiment. We report here the relationship of serpin's conformational switch mechanism with Inhibitory activity. There is little direct correlation between loop insertion rate and inhibitory activity. Rather, disrupting a salt bridge between R196 and E354 accelerates loop translocation even though it impairs the inhibitory activity. Moreover, the serpin's reactive site loop is translocated, at least partially, prior to loop cleavage.

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농어촌(農漁村)과 시설거주(施設居住) 아동(兒童)의 영양상태(營養狀態)에 관(關)한 연구(硏究) -제2보(第2報) : 혈액성장(血液性狀)을 중심(中心)으로- (A Study on the Nutritional Status of Children Living in Rural Area and City Orphan Asylum -II. Hematological view-)

  • 박복희
    • 한국식품영양과학회지
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    • 제14권2호
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    • pp.108-116
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    • 1985
  • This study was designed to investigate the nutritional anemic state and hematological findings of children, aged 11 and 12 years, living in Haenam Koon (living at home) and in Mokpo City (living in an orphan asylum) located in Chonnam. The mean red blood cell number of male was higher than female in both groups of living in the rural area and in the city orphan asylum (p<0.01), but the white blood cell count was not significantly different. The levels of average hemoglobin, hematocrit and mean corpuscular hemoglobin concentration of the children in both groups were similar, and 19.8% of children living at home in the rural area and 32.1% of children living in the city orphan asylum were anemic. The mean levels of serum total protein, albumin and A/G ratio in children of both group were not statistically different, and 18.4% of children living at home in the rural area and 13.2% of children living in the city orphan asylum were insufficient in the serum total protein value. The average serum cholesterol level of children living at home in the rural area was higher than that of children living in the city orphan asylum(p<0.01), and that of female living at home was higher than that of male (p<0.05). The mean levels of serum iron, total iron binding capacity and transferrin saturation of children living at home in the rural area were significantly higher than those of children living in the city orphan asylum (p<0.01).

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Acetylcholinesterase(AChE)-Catalyzed Hydrolysis of Long-Chain Thiocholine Esters: Shift to a New Chemical Mechanism

  • Jung, Dai-Il;Shin, Young-Ju;Lee, Eun-Seok;Moon, Tae-sung;Yoon, Chang-No;Lee, Bong-Ho
    • Bulletin of the Korean Chemical Society
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    • 제24권1호
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    • pp.65-69
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    • 2003
  • The kinetic and chemical mechanisms of AChE-catalyzed hydrolysis of short-chain thiocholine esters are relatively well documented. Up to propanoylthiocholine (PrTCh) the chemical mechanism is general acid-base catalysis by the active site catalytic triad. The chemical mechanism for the enzyme-catalyzed butyrylthiocholine(BuTCh) hydrolysis shifts to a parallel mechanism in which general base catalysis by E199 of direct water attack to the carbonyl carbon of the substrate. [Selwood, T., et al. J. Am. Chem. Soc. 1993, 115, 10477- 10482] The long chain thiocholine esters such as hexanoylthiocholine (HexTCh), heptanoylthiocholine (HepTCh), and octanoylthiocholine (OcTCh) are hydrolyzed by electric eel acetylcholinesterase (AChE). The kinetic parameters are determined to show that these compounds have a lower Michaelis constant than BuTCh and the pH-rate profile showed that the mechanism is similar to that of BuTCh hydrolysis. The solvent isotope effect and proton inventory of AChE-catalyzed hydrolysis of HexTCh showed that one proton transfer is involved in the transition state of the acylation stage. The relationship between the dipole moment and the Michaelis constant of the long chain thiocholine esters showed that the dipole moment is the most important factor for the binding of a substrate to the enzyme active site.

수산업에 대한 국가 역할: 헌법의 정치경제학적 의미와 해석 (National Roles of Fisheries: Political-Economic Meaning and Interpretation of the Constitution)

  • 박성쾌
    • 수산해양교육연구
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    • 제18권3호
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    • pp.244-260
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    • 2006
  • The main objective of this study aims at analyzing the national roles of fisheries in the global economy from the constitutional view point. Globalization provides general firms and businessmen with great opportunities that allow them to be able to plan and operate their business strategies beyond the national boundaries. WTO and FTA negotiations must be important facilitators of such globalization. However, Korean primary industries like fisheries under comparative disadvantage have a high probability that there may have to be the trade-offs between the primary and the industrial sector to maximize national benefits in the process of bilateral and/or multilateral international trade deals. Since, moreover, fishermen face strong binding constraints under which they hardly carry out fishing operations standing aloof from national boarders, they have to manage fishing businesses within their own national territory, to maintain and develop fisheries culture, and to explore their own destiny for themselves. Because of such reasons, the constitution imposes upon the State the duty to support and develop fisheries and fishing villages. Considering its article and spirit associated with fisheries(i.e. article 123), it seems unnecessary at this point in time that Korean society makes debates over national consensus for supporting fisheries. The reason is because the explicit provisions of the constitution do not allow their arbitrary interpretation depending upon interests or policy situations. However, where national support to the particular sector could not meet the expected social value system, there would be a great deal of chance to invite serious societal debates over such national commitment to the fisheries. Therefore, whether using it efficiently and realizing the socially expected policy goals must be a responsibility of both fisheries administration and fishing industries.

사염화탄소와 갈락토사민 간장해 시의 오큐빈의 체내동태 차이 (Different Pharmacokinetics of Aucubin in Rats of Carbon tetrachloride and D-Galactosamine-induced Hepatic Failure)

  • 김미형;심창구;장일무
    • 약학회지
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    • 제37권4호
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    • pp.383-388
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    • 1993
  • Pharmacokinetics of aucubin, an irdoid glucoside, was compared in rats of experimental hepatic failure(EHF). EHF was induced by CCI$_{4}$ or D-galactosamine pretreatment. This work was designed to find out any differences in the pharmacokinetics of aucubin that may explain the different protective effect of aucubin on CCI$_{4}$- and galactosamine-induced EHF : aucubin reportedly protected CCI$_{4}$-inducing hepatotoxicity effectively, but did not for galactosamine-hepatotoxicity. EHF was induced by intraperitoneal injection Of CCI$_{4}$(0.9ml/kg) or galactosamine(250 mg/kg) to Wistar rats 24 hr before the pharmacokinetic study. The rats were fasted during the 24 hr. Aucubin was iv injected at a dose of 15 mg/kg and the plasma aucubin was assayed by HPLC. There were no significant differences in the pathophysiologies(body weight, liver weight, GTP, hematocrit, blood cell distrbution and plasma protein binding of aucubin) between the two EHF models except GOP which was significantly (p<0.05) higher in CCI$_{4}$-than in galactosamine-EHF. On the other hand, pharmacokinetics of aucubin such as total cleatance(CL$_{t}$), distribution volume at steady-state(Vd$_{ss}$), and mean residence time(MRT) differed significantly(p<0.05) between the models : for example, CL$_{t}$ was increased two fold by CCI$_{4}$, but not by galaclosamine ; Vd$_{ss}$, in galactosamine-EHF was higher than that in CCI$_{4}$-EHF ; MRT was decreased by CCI$_{4}$, but increased conversely by galactosamine. The increase of CL$_{t}$(and decrease of MRT) in rats of CCI$_{4}$-EHF was contrary to the general expectation for the hepatic failure : most of the hepatic failures have been known to decrease CL$_{t}$ of the administered drugs. Whether the difference in the pharmacokinetics is responsible for the different protective effect of aucubin against the two EHF models is of interest. However, much more studies on biliary excretion, urinary excretion, and hepatic uptake in cellular level should be preceded before any conclusions are made on the role of different pharmacokinetics on the different pharmacology of aucubin.

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Structure of the Tripartite Multidrug Efflux Pump AcrAB-TolC Suggests an Alternative Assembly Mode

  • Kim, Jin-Sik;Jeong, Hyeongseop;Song, Saemee;Kim, Hye-Yeon;Lee, Kangseok;Hyun, Jaekyung;Ha, Nam-Chul
    • Molecules and Cells
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    • 제38권2호
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    • pp.180-186
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    • 2015
  • Escherichia coli AcrAB-TolC is a multidrug efflux pump that expels a wide range of toxic substrates. The dynamic nature of the binding or low affinity between the components has impeded elucidation of how the three components assemble in the functional state. Here, we created fusion proteins composed of AcrB, a transmembrane linker, and two copies of AcrA. The fusion protein exhibited acridine pumping activity, suggesting that the protein reflects the functional structure in vivo. To discern the assembling mode with TolC, the AcrBA fusion protein was incubated with TolC or a chimeric protein containing the TolC aperture tip region. Three-dimensional structures of the complex proteins were determined through transmission electron microscopy. The overall structure exemplifies the adaptor bridging model, wherein the funnel-like AcrA hexamer forms an intermeshing cogwheel interaction with the ${\alpha}$-barrel tip region of TolC, and a direct interaction between AcrB and TolC is not allowed. These observations provide a structural blueprint for understanding multidrug resistance in pathogenic Gram-negative bacteria.

Open channel block of Kv1.4 potassium channels by aripiprazole

  • Park, Jeaneun;Cho, Kwang-Hyun;Lee, Hong Joon;Choi, Jin-Sung;Rhie, Duck-Joo
    • The Korean Journal of Physiology and Pharmacology
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    • 제24권6호
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    • pp.545-553
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    • 2020
  • Aripiprazole is a quinolinone derivative approved as an atypical antipsychotic drug for the treatment of schizophrenia and bipolar disorder. It acts as with partial agonist activities at the dopamine D2 receptors. Although it is known to be relatively safe for patients with cardiac ailments, less is known about the effect of aripiprazole on voltage-gated ion channels such as transient A-type K+ channels, which are important for the repolarization of cardiac and neuronal action potentials. Here, we investigated the effects of aripiprazole on Kv1.4 currents expressed in HEK293 cells using a whole-cell patch-clamp technique. Aripiprazole blocked Kv1.4 channels in a concentration-dependent manner with an IC50 value of 4.4 μM and a Hill coefficient of 2.5. Aripiprazole also accelerated the activation (time-to-peak) and inactivation kinetics. Aripiprazole induced a voltage-dependent (δ = 0.17) inhibition, which was use-dependent with successive pulses on Kv1.4 currents without altering the time course of recovery from inactivation. Dehydroaripiprazole, an active metabolite of aripiprazole, inhibited Kv1.4 with an IC50 value of 6.3 μM (p < 0.05 compared with aripiprazole) with a Hill coefficient of 2.0. Furthermore, aripiprazole inhibited Kv4.3 currents to a similar extent in a concentration-dependent manner with an IC50 value of 4.9 μM and a Hill coefficient of 2.3. Thus, our results indicate that aripiprazole blocked Kv1.4 by preferentially binding to the open state of the channels.