• Title/Summary/Keyword: Substrate interaction

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Parkin Interacts with the PDZ Domain of Multi-PDZ Domain Protein MUPP1 (Parkin과 Multi-PDZ Domain Protein (MUPP1) 단백질 간의 PDZ 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Lee, Won Hee;Kim, Mooseong;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.8
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    • pp.820-826
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    • 2014
  • The localization to specific subcellular sites and the regulation of cell surface receptors and channels are crucial for proper functioning. Postsynaptic density-95/Disks large/Zonula occludens-1 (PDZ)-domain is involved in recognition of and interaction between various proteins, by which the localization and the regulation are mediated. Multi-PDZ domain protein 1 (MUPP1) contains 13 PDZ domains. MUPP1 serves a scaffolding function for structure proteins and signaling proteins, but the mechanism how MUPP1 is stabilized and signalized has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and Parkin. Parkin is an E3 ubiquitin ligase. Loss-of-function mutations of Parkin gene are known to cause an autosomal recessive juvenile parkinsonism. Parkin bound to the $12^{th}$ PDZ domain, but not to other PDZ domains of MUPP1. The C-terminal end of Parkin has a type II PDZ-association motif, which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. When co-expressed in HEK-293T cells, Parkin co-localized with MUPP1. When co-expressed with ubiquitin in HEK-293T cells, MUPP1 has been strongly ubiquitinated by Parkin. These findings collectively suggest that MUPP1 is a novel substrate of Parkin and its function or stability could be modulated by Parkin-mediated ubiquitination.

Effect of Nitric Oxide on the Sinusoidal Uptake of Organic Cations and Anions by Isolated Hepatocytes

  • Song, Im-Sook;Lee, In-Kyoung;Chung, Suk-Jae;Kim, Sang-Geon;Lee, Myung-Gull;Shim, Chang-Koo
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.984-988
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    • 2002
  • The issue of whether or not the presence NOx (NO and oxidized metabolites) in the hepatocytes at pathological levels affects the functional activity of transport systems within the sinusoidal membrane was investigated. For this purpose, the effect of the pretreatment of isolated hepatocytes with sodium nitroprusside (SNP), a spontaneous NO donor, on the sinusoidal uptake of tributylmethylammonium (TBuMA) and triethylmethyl ammonium (TEMA), representative substrates of the organic cation transporter (OCT), and taurocholate, a representative substrate of the $Na^+$/taurocholate cotransporting polypeptide (NTCP), was measured. The uptake of TBuMA and TEMA was not affected by the pretreatment, as demonstrated by the nearly identical kinetic parameters for the uptake ($i.e., V_{max}, K_{m} and CL_{linear}$). The uptake of mannitol into hepatocytes was not affected, demonstrating that the membrane integrity remained constant, irregardless of the SNP prutreatment. On the contrary, the uptake of taurocholate was significantly inhibited by the pretreatment, resulting in a significant decrease in V_{max}$, thus providing a clear demonstration that NOx preferentially affects the function of NTCP rather than OCT on the sinusoidal membrane. A direct interaction between NOx and NTCP or a decrease in $Na^+/K^+$ ATPase activity as the result of SNP pretreatment might be responsible for this selective effect of NOx.

Enhanced Bone-Regenerative Performance of Porous Hybrid Scaffolds by Surface Immobilization of Nano-Hydroxyapatite

  • Lee, Sang-Cheon
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.11a
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    • pp.12.1-12.1
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    • 2009
  • Nano-hydroxyapatite (N-HAp)has shown the pivotal role in producing bone-regenerative materials since it has similarity to natural bone minerals in terms of size, morphology, and the composition. Currently, the combination of biopolymers and N-HAp is recognizedas an attractive approach in generating hybrid scaffolds for bone tissueengineering. Surface engineering is an important issue since it determines whether cells can effectively adhere and proliferate on porous scaffolds. We aim to develop a synthetic approach to porous 3D scaffolds by immobilizing N-HAp on pore surfaces. The discrete nano-level anchoring of N-HAp on the scaffold pore surface is achieved using surface-repellent stable colloidal N-HAp with surface phosphate functionality. This rational surface engineering enables surface-anchored N-HAp to express its overall intrinsic bioactivity,since N-HAp is not phase-mixed with the polymers. The porous polymer scaffolds with surface-immobilized N-HAp provide more favorable environments thanconventional bulk phase-mixed polymer/N-HAp scaffolds in terms of cellular interaction and growth. In vitro biological evaluation using alkalinephosphatase activity assay supports that immobilized N-HAp on pore surfaces of polymer scaffolds contributed to the more enhanced in vitro osteogenicpotential. Besides, the scaffolds with surface-exposed N-HAp provide favorable environments for enhanced in vivo bone tissue growth, estimated by characteristic biomarkers of bone formation such as collagen. The results suggest that newly developed hybrid scaffolds with surface-immobilized N-HApmay serve as a useful 3D substrate with pore surfaces featuring excellent bonetissue-regenerative properties. Acknowledgement. This research was supported by a grant (code #: 2009K000430) from 'Center for Nanostructured Materials Technology' under '21st Century Frontier R&D Programs' of the Ministry of Education, Science and Technology, Korea.

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Electrochemical Detection of $17{\beta}-estradiol$ by using DNA Aptamer Immobilized Nanowell Gold Electrodes

  • Kim, Yeon-Seok;Jung, Ho-Sup;Lee, Hea-Yeon;Kawai, Tomoji;Gu, Man-Bock
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.88-92
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    • 2005
  • Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

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CD30-Mediated Regulation of Cell Adhesion Molecule Expression on Murine T Cells

  • Nam, Sang-Yun
    • IMMUNE NETWORK
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    • v.3 no.1
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    • pp.8-15
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    • 2003
  • Background: CD30 is a member of TNF receptor family and expressed on lymphocytes and other hematopoietic cells following activation as well as Hodgkin and Reed-Sternberg cells in Hodgkin's lymphoma. In this study, CD30-mediated regulation of cell adhesion molecule expression on normal activated mouse T cells was investigated. Methods: Mouse T cells were activated with anti-CD3 antibody for induction of CD30, which was cross-linked by immobilized anti-CD30 antibody. Results: High level of CD30 expression on T cells was observed on day 5, but only little on day 3 even under culture condition resulting in an identical T cell proliferation, indicating that CD30 expression requires a prolonged stimulation up to 5 days. Cross-linking of CD30 alone altered neither proliferation nor apoptosis of normal activated T cells. Instead, CD30 appeared to promote cell adherence to culture substrate, and considerably upregulated ICAM-1 and, to a lesser extent, ICAM-2 expression on activated T cells, whereas CD2 and CD18 (LFA-1) expression was not affected. None of cytokines known as main regulators of ICAM-1 expression on tissue cells (IL 4, $IFN{\gamma}$ and $IFN{\alpha}$) enhanced ICAM-1 expression in the absence of CD30 signals. On the other hand, addition of $NF-{\kappa}B$ inhibitor, PDTC (0.1 mM) completely abrogated the CD30-mediated upregulation of ICAM-1 expression, but not CD2 and ICAM-2 expression. Conclusion: This results support that CD30 upregulates ICAM-1 expression of T cell and such regulation is not mediated by higher cytokine production but $NF-{\kappa}B$ activation. Therefore, CD30 may play important roles in T-T or T-B cell interaction through regulation of ICAM-1, and -2 expression.

Encapsulation and optical properties of Er3+ ions for planar optical amplifiers via sol-gel process (졸-겔법을 이용한 광증폭기의 Er 이온 캡슐화 및 광학적 특성)

  • Kim, Joo-Hyeun;Seok, Sang-Il;Ahn, Bok-Yeop
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2003.11a
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    • pp.135-135
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    • 2003
  • The fast evolution in the fold of optical communication systems demands powerful optical information treatment. These functions can be performed by integrated optical systems. A key component of such systems is erbium doped waveguide amplifier(EDWA). The intra 4f radiative transition of Er at 1.5 $\mu\textrm{m}$ is particularly interesting because this wavelength is standard in optical telecommunications. The fabrication of waveguide amplifier for integrated optics using sol-gel process has received an increasing attention. Potential advantage of lower cost by less capital equipment and easy processing makes this process an attractive alternatives to conventional technologies like flame hydrolysis deposition, ion exchange and chemical vapor deposition, etc. In addition, sol-gel process has been found to be extremely suitable for the control of composition and refractive index related directly with optical properties. The main drawback of such an amplifier with respect to the EDWA is the need for a much higher Er3+ concentration to compensate for the smaller interaction length. However, the high doping of Er might be resulted in the non-radiative relaxation by clustering of Er ions End co-operative upconversion. In order to solve this problem, we investigate the possibility of avoiding short Er-Er distances by encapsulation of Er3+ ions in hosts such as organic-inorganic hybrid materials. For inorganic-organic hybrid sols, methacryloxypropyltrimethoxysilane (MPTS), zirconyl chloride octahydrate and erbium(III) chloride hexahydrate were used as starting materials, followed by conventional sol-gel process. It was observed by TEM that nano sols having core/shell toplology were formed, depending on the mole ratio of Zr/Er. The surface roughness for the coatings on Si substrate was investigated by AFM as a function of Zr/Er ratio. The local environment and vibrational Properties of Er3+ ions were studied using Near-IR, FT-IR, and UV/Vis spectroscopy. Nano hybrid coatings derived from polymer and Er doped encapsulation Eave the good luminescence at 1.55$\mu\textrm{m}$.

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Binding of Lichen Phenolics to Purified Secreted Arginase from the Lichen Evernia prunastri

  • Legaz, Maria-Estrella;Vicente, Carlos;Pedrosa, Mercedes M.
    • BMB Reports
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    • v.34 no.3
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    • pp.194-200
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    • 2001
  • Secreted arginase from Evernia prunastri thallus has been purified 616-fold from the incubation medium. Purified arginase was resolved as only one peak in a capillary electrophoresis with a pI value of 5.35. The protein contained high amounts of acidic amino acids, such as Asx and Glx, and a relatively high quantity of Ser and Gly. The molecular mass of native, purified arginase was estimated as about 26 kDa by SE-HPLC. Substrate saturated kinetic showed a typical Michaelis-Menten relationship with a K_m value of 3.3 mM L-arginine. Atranorin behaved as a mixed activator of the enzyme (apparent $K_m$ = 0.96 mM); whereas evernic and usnic acid were revealed as non competitive inhibitors (apparent $K_m$ values were 3.16 mM and 3.05 mM, respectively). Kinetics of atranorin binding indicated that saturation was reached from 0.18 ${\mu}mol$ of the total atranorin and the occurrence of multiple sites for the ligand. This agrees with a possible aggregation of several enzyme subunits during the interaction process. A value of binding sites of about 12 was obtained. The binding of evernic acid was saturated from 23 nmol of total phenol. The number of binding sites was about 5. The loss of the binding ability of evernic acid could be interpreted as a single negative cooperatively. Usnic acid behaves in a similar way to evernic acid, although the binding saturation occurs at $0.14\;{\mu}moles$ of the ligand. This binding appears to be unspecific, and has 28 usnic acid binding sites to the protein.

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Support Effect of Arc Plasma Deposited Pt Nanoparticles/TiO2 Substrate on Catalytic Activity of CO Oxidation

  • Qadir, Kamran;Kim, Sang Hoon;Kim, Sun Mi;Ha, Heonphil;Park, Jeong Young
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.261-261
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    • 2013
  • The smart design of nanocatalysts can improve the catalytic activity of transition metals on reducible oxide supports, such as titania, via strong metal-support interactions. In this work, we investigatedtwo-dimensional Pt nanoparticle/titania catalytic systems under the CO oxidation reaction. Arc plasma deposition (APD) and metal impregnation techniques were employed to achieve Pt nanoparticle deposition on titania supports, which were prepared by multitarget sputtering and sol-gel techniques. APD Pt nanoparticles with an average size of 2.7 nm were deposited on sputtered and sol-gel-prepared titania films to assess the role of the titania support on the catalytic activity of Pt under CO oxidation. In order to study the nature of the dispersed metallic phase and its effect on the activity of the catalytic CO oxidation reaction, Pt nanoparticles were deposited in varying surface coverages on sputtered titania films using arc plasma deposition. Our results show an enhanced activity of Pt nanoparticles when the nanoparticle/titania interfaces are exposed. APD Pt shows superior catalytic activity under CO oxidation, as compared to impregnated Pt nanoparticles, due to the catalytically active nature of the mild surface oxidation and the active Pt metal, suggesting that APD can be used for large-scale synthesis of active metal nanocatalysts.

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Design and Implementation of CPW-Fed UWB Monopole Antenna (CPW 급전 방식을 이용한 UWB 모노폴 안테나 설계 및 구현)

  • Yu, Ju-Bong;Jeon, Jun-Ho;An, Chan-Kyu;Kim, Woo-Chan;Yang, Woon-Geun
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.21 no.2
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    • pp.218-223
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    • 2010
  • In this paper, a novel CPW(Coplanar Waveguide)-fed UWB(Ultra Wide Band) antenna is designed, implemented, and measured for UWB communications. CPW-fed planar antenna has advantages of wide-bandwidth, low-cost and easy interaction with radio frequency front end circuitry. We have used HFSS(High Frequency Structure Simulator) of Ansoft which is based on the FEM(Finite Element Method) to simulate the proposed antenna. FR-4 substrate of thickness 1.6 mm and relative permitivity 4.4 is used for implementation. The proposed antenna showed VSWR(Voltage Standarding Wave Ratio)${\leq}2$ for the frequency band from 3.1 GHz to 10.6 GHz which is used for ultra wide band communication. Measured peak gains are 2.61, 4.95, 2.89, 7.35 dBi at 3, 6, 8, 11 GHz, respectively.

Fabrication of carbon nanotube emitters by filtration through a metal mesh

  • Choi, Ju-Sung;Lee, Han-Sung;Gwak, Jeung-Chun;Lee, Nae-Sung
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2010.06a
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    • pp.150-150
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    • 2010
  • Carbon nanotubes have drawn attention as one of the most promising emitter materials ever known not only due to their nanometer-scale radius of curvature at tip and extremely high aspect ratios but also due to their strong mechanical strength, excellent thermal conductivity, good chemical stability, etc. Some applications of CNTs as emitters, such as X-ray tubes and microwave amplifiers, require high current emission over a small emitter area. The field emission for high current density often damages CNT emitters by Joule heating, field evaporation, or electrostatic interaction. In order to endure the high current density emission, CNT emitters should be optimally fabricated in terms of material properties and morphological aspects: highly crystalline CNT materials, low gas emission during electron emission in vacuum, optimal emitter distribution density, optimal aspect ratio of emitters, uniform emitter height, strong emitter adhesion onto a substrate, etc. We attempted a novel approach to fabricate CNT emitters to meet some of requirements described above, including highly crystalline CNT materials, low gas emission, and strong emitter adhesion. In this study, CNT emitters were fabricated by filtrating an aqueous suspension of highly crystalline thin multiwalled CNTs (Hanwha Nanotech Inc.) through a metal mesh. The metal mesh served as a support and fixture frame of CNT emitters. When 5 ml of the CNT suspension was engaged in filtration through a 400 mesh, the CNT layers were formed to be as thick as the mesh at the mesh openings. The CNT emitter sample of $1{\times}1\;cm^2$ in size was characteristic of the turn-on electrical field of 2.7 V/${\mu}m$ and the current density of 14.5 mA at 5.8 V/${\mu}m$ without noticeable deterioration of emitters. This study seems to provide a novel fabrication route to simply produce small-size CNT emitters for high current emission with reliability.

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