• Title/Summary/Keyword: SH3

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Establishment of a Binding Assay System for Screening of the Inhibitors of $p56^{lck}$ SH2 Domain

  • Kim, Jyn-Ho;Hur, Eun-Mi;Yun, Yung-Dae
    • BMB Reports
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    • v.31 no.4
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    • pp.370-376
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    • 1998
  • Src-Homology 2 (SH2) domains have a capacity to bind phosphotyrosine-containing sequence context and play essential roles in various cellular signaling pathways. Due to the specific nature of the binding between SH2 domains and their counterpart proteins, inhibitors of SID domain binding have drawn extensive attention as a potential candidate for therapeutic agents. Here, we describe the binding assay system to screen for the ligands or blockers of the SH2 domains with an emphasis on the $p56^{lck}$ SH2 domain. In our assay system, SID domains expressed and purified as fusion proteins to Glutathione-S-transferase (GST) were covalently attached to 96-well microtitre plates through amide bond formation, which were subsequently allowed to bind the biotinylated phosphotyrosine (pY)containing synthetic pep tides. The binding of biotinylated pY peptides was detected by the horseradish peroxidase (HRP)-conjugated streptavidin. Using the various combinations of SH2 domain-pY peptides, we observed that: (1) The binding of pY-peptides to its counterpart SH2 domain is concentration-dependent and saturable; (2) The binding is highly specific for a particular combination of SH2 domain-pY peptide pair; and (3) The binding of Lck SH2-cognate pY-peptides is specifically competed by the nonbiotinylated peptides with expected relative affinity. These results indicate that the established assay system detects the SH2-pY peptide interaction with reproducible sensitivity and specificity and is suitable for screening the specific inhibitors of $p56^{lck}$ SH2 function.

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EFFICIENCY OF ENERGY TRANSFER BY A POPULATION OF THE FARMED PACIFIC OYSTER, CRASSOSTREA GIGAS IN GEOJE-HANSAN BAY (거제${\cdot}$한산만 양식굴 Crassostrea gigas의 에너지 전환 효율)

  • KIM Yong Sool
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.13 no.4
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    • pp.179-183
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    • 1980
  • The efficiency of energy transfer by a population of the farmed pacific oyster, Crassostrea gigas was studied during culture period of 10 months July 1979-April 1980, in Geoje-Hansan Bay near Chungmu City. Energy use by the farmed oyster population was calculated from estimates of half-a-month unit age specific natural mortality rate and data on growth, gonad output, shell organic matter production and respiration. Total mortality during the culture period was estimated approximate $36\%$ from data on survivor individual number per cluster. Growth may be dual consisted of a curved line during the first half culture period (July-November) and a linear line in the later half period (December-April). The first half growth was approximated by the von Bertalanffy growth model; shell height, $SH=6.33\;(1-e^{0.2421(t+0.54)})$, where t is age in half-a-month unit. In the later half growth period shell height was related to t by SH=4.44+0.14t. Dry meat weight (DW) was related to shell height by log $DW=-2.2907+2.589{\cdot}log\;SH,\;(2, and/or log $DW=-5.8153+7.208{\cdot}log\;SH,\;(5. Size specific gonad output (G) as calculated by condition index of before and after the spawning season, was related to shell height by $G=0.0145+(3.95\times10^{-3}{\times}SH^{2.9861})$. Shell organic matter production (SO) was related to shell height by log $SO=-3.1884+2.527{\cdot}1og\;SH$. Size and temperature specific respiration rate (R) as determined in biotron system with controlled temperature, was related to dry meat weight and temperature (T) by log $R=(0.386T-0.5381)+(0.6409-0.0083T){\cdot}log\;DW$. The energy used in metabolism was calculated from size, temperature specific respiration and data on body composition. The calorie contents of oyster meat were estimated by bomb calorimetry based on nitrogen correction. The assimilation efficiency of the oyster estimated directly by a insoluble crude silicate method gave $55.5\%$. From the information presently available by other workers, the assimilation efficiency ranges between $40\%\;and\;70\%$. Twenty seven point four percent of the filtered food material expressed by energy value for oyster population was estimated to have been rejected as pseudofaeces : $17.2\%$ was passed as faeces; $35.04\%$ was respired and lost as heat; $0.38\%$ was bounded up in shell organics; $2.74\%$ was released as gonad output, $2.06\%$ was fell as meat reducing by mortality. The remaining $15.28\%$ was used as meat production. The net efficiency of energy transfer from assimilation to meat production (yield/assimilation) of a farm population of the oyster was estimated to be $28\%$ during culture period July 1979-April 1980. The gross efficiency of energy transfer from ingestion to meat production (yield/food filtered) is probably between $11\%\;and\;20\%$.

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The Effect of Salicylic Acid on the Accumulation of $Cd^{2+}$ and Nonprotein-SH Synthesis in Roots and Epidermal Strips of Commelina Communis L. (닭의장풀에 $Cd^{2+}$처리 시 $Cd^{2+}$ 흡수와 nonprotein-SH 합성에 미치는 살리실릭산의 영향)

  • 이준상
    • Korean Journal of Environmental Biology
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    • v.19 no.3
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    • pp.218-222
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    • 2001
  • Three weeks old Commelina was cultivated in Hoagland solution with or without $(\pm{100\mu{M}\;Cd^{2+}$ and with $100\mu{M}\;Cd^{2+}+100\mu{M\; SA)$ for 4 days. The contents of $Cd^{2+}$ and nonprotein-SH synthesis in roots and isolated epidermal strips were measured. In the treatment of $Cd^{2+}$, the concentration of $Cd^{2+}$ was $98\mu{M}\;Cd^{2+}$/g.fr.wt and it was 6 times higher than that of control. $Cd^{2+}$+SA increased 1.2 times higher concentration of $Cd^{2+}$ than that of the treatment ofTEX>$Cd^{2+}$ alone. In the treatment of TEX>$Cd^{2+}$, nonprotein-SH also increased 2 times higher than that of the control. TEX>$Cd^{2+}$ + SA increased 2 times higher concentration of nonprotein-SH than that of the treatment of TEX>$Cd^{2+}$ as well. In case of isolated epidermal strips, the response was similar with the roots. Therefore, it suggest that the effect of salicylic acid on the accumulation of TEX>$Cd^{2+}$ and nonprotein-SH synthesis in roots, and isolated epidermal strips was related with an increase of the membrane permeability by SA and that SA may be positively involved in the process of nonprotein- SH synthesis.

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Transcriptional Regulation of Human GD3 Synthase (hST8Sia I) by Fenretinide in Human Neuroblastoma SH-SY-5Y Cells (사람 신경모세포종 세포주 SH-SY5Y에서 fenretinide에 의한 GD3합성효소(hST8Sia I)의 전사조절기작)

  • Kang, Nam-Young;Kwon, Haw-Young;Lee, Young-Choon
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1332-1338
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    • 2010
  • To elucidate the mechanism underlying the regulation of hST8Sia I gene expression in FenR-induced SH-SY5Y cells, we characterized the promoter region of the hST8Sia I gene. Functional analysis of the 5'-flanking region of the hST8Sia I gene showed that the -1146 to -646 region functions as the FenR-inducible promoter of hST8Sia I in SH-SY5Y cells. Site-directed mutagenesis indicated that the NF-&B binding site at -731 to -722 was crucial for the FenR-induced expression of hST8Sia I in SH-SY5Y cells. To investigate which signal transduction pathway was involved in FenR-stimulated induction of hST8Sia I in SH-SY5Y cells, we performed Western blot analysis using phospho-specific antibodies in order to measure their degree of regulatory phosphorylation. Phosphorylations of AKT and RelA (p65) subunit of NF-${\kappa}B$ were significantly elevated in cytosolic and nuclear fractions of FenR-stimulated SH-SY5Y cells, respectively, than in control or DMSO-treated SH-SY5Y cells. These results suggest that FenR induce transcriptional up-regulation of hST8Sia I gene expression through translocation of RelA (p65) subunit of NF-${\kappa}B$ to nucleus by AKT signal pathway in SH-SY5Y cells.

Design and Implementation of Matching Engine for QbSH System Based on Polyphonic Music (다성음원 기반 QbSH 시스템을 위한 매칭엔진의 설계 및 구현)

  • Park, Sung-Joo;Chung, Kwang-Sue
    • Journal of Korea Multimedia Society
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    • v.15 no.1
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    • pp.18-31
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    • 2012
  • This paper proposes a matching engine of query-by-singing/humming (QbSH) system which retrieves the most similar music information by comparing the input data with the extracted feature information from polyphonic music like MP3. The feature sequences transcribed from polyphonic music may have many errors. So, to reduce the influence of errors and improve the performance, the chroma-scale representation, compensation and asymmetric DTW (Dynamic Time Warping) are adopted in the matching engine. The performance of various distance metrics are also investigated in this paper. In our experiment, the proposed QbSH system achieves MRR (Mean Reciprocal Rank) of 0.718 for 1000 singing/humming queries when searching from a database of 450 polyphonic musics.

The Molecular Simulation Study for the Adsorption of $H_2S,\;NH_3$ and $CH_3SH$ on Graphite Carbon (Graphite Carbon에 $H_2S,\;NH_3$$CH_3SH$의 흡착에 대한 분자모사 연구)

  • 신창호;김종열;이영택;김정열;김승준
    • Journal of the Korean Society of Tobacco Science
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    • v.25 no.1
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    • pp.59-69
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    • 2003
  • The adsorption characteristics of H$_2$S, NH$_3$and $CH_3$SH on the graphite carbon have been investigated using Grand Canonical Monte Carlo(GCMC) method with universal force field (UFF) and dreiding force field. Most of the activated carbons used in vapor phase adsorption have the micropore of 6$\AA$ to 20$\AA$ and the specific surface area of ca. 1000 m$^2$/g, as the result of $N_2$ adsorption by BET method. For the more efficient comparison, the activated carbons have been manipulated with different pore sizes. The adsorption characteristics of H$_2$S, NH$_3$and $CH_3$SH have been considered at various temperatures and pressures. The adsorption amount using Dreiding force field is predicted to be lower than that using UFF. As the temperature is going to high, the adsorption amount of adsorbates is decreased due to their vaporization. Considering the pore size effect, the adsorption characteristic depends on the adsorbate size, polarity and interaction between adsorbates, etc. At all cases employed in this study, NH$_3$ is barely adsorbed and $CH_3$SH is preferentially adsorbed on the graphite carbon. Our theoretical result is qualitatively good agreement with the experimental observation. However, there are some quantitative discrepancies depending on the functional groups and pore size distribution on the real activated carbons used in experiment.

The Effector Functions of Mature T Lymphocytes Are Impaired in Transgenic Mice Expressing the SH2 Domain of TSAd/Lad

  • Choi, Youngbong;Park, Eunkyung;Ahn, Eunseon;Park, Inyoung;Yun, Yungdae
    • Molecules and Cells
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    • v.28 no.3
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    • pp.183-188
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    • 2009
  • TSAd/Lad is a T cell adaptor molecule involved in $p56^{lck}$-mediated T cell activation. To investigate the functions of TSAd in T cells, we generated transgenic (TG) mice expressing the SH2 domain of TSAd (TSAd-SH2) under the control of the $p56^{lck}$ proximal promoter. In T cells from TSAd-SH2 TG mice, T cell receptor (TCR)-mediated early signaling events, such as $Ca^{2+}$ flux and ERK activation, were normal; however, late activation events, such as IL-2 production and proliferation, were significantly reduced. Moreover, TCR-induced cell adhesion to extracellular matrix (ECM) proteins and migration through ECM proteins were defective in T cells from TSAd-SH2 TG mice. Furthermore, the contact hypersensitivity (CHS) reaction, an inflammatory response mainly mediated by T helper 1 (Th1) cells, was inhibited in TSAd-SH2 TG mice. Taken together, these results show that TSAd, particularly the SH2 domain of TSAd, is essential for the effector functions of T cells.

Effects of mold temperature on the part dimension and surface quality of the injection molded cavity filter (금형온도가 Cavity Filter 성형품의 치수 및 외관품질에 미치는 영향에 관한 연구)

  • 김동학;김태완
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.4 no.3
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    • pp.164-167
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    • 2003
  • In this study, we developed the mold for the plastic cavity filter which is a kind of a wireless communication device. Since the cavity filter is made of aluminium, the cost of fabrication is high and the production rate is low. But we can produce plastic cavity filter part by injection molding process with the mold which was designed by our team. The dimension and surface quality of plastic cavity filter was investigated by varying the molding method (conventional and MmSH process) and two different types of resin(PC/ABS and ABS). In case of ABS part, the shrinkage of the inner partition walls was decreased when we adopted MmSH method. The weight of both ABS and PC/ABS parts increased and the surface roughness decreased with MmSH process.

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Experimental Study on Condensation Heat Transfer Characteristics of Special Heat Transfer Tubes (응축용 특수 전열관의 열전달 특성에 관한 연구)

  • 한규일;박종운;권영철;조동현
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.13 no.9
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    • pp.827-835
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    • 2001
  • In this study, condensation heat transfer characteristics were conducted with special heat transfer tubes of SH-C type. Experiments were carried out the saturated vapor temperature of 334K and the wall subcooling of 1.5-4.5K. The refrigerant was R-113 and the enhanced tubes used in the present study were SH-CDR, SH-CYR and SH-CHR. The experimental results showed that the condensation heat transfer coefficients of SH-C type tubes were about 23-66% higher than those of a low integral-fin tube. It was visualized that the condensed liquid on the outer surface of SH-C type tubes flowed continuously down unlike a low integral-fin tube and a plain tube, due to a 3-D extending fin on the outer surface of SH-C type tubes. As a result, the thermal resistance of the condensed liquid decreased and the heat transfer coefficient increased. Also, the enhancement ratio of SH-CDR tube was the highest, and it was about 9-11 times as compared to that of a plain tube.

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Study on barrier characteristics of STM tip/Viologen molecules and morphology (STM tip/Viologen 분자의 Barrier특성과 모폴로지 촉정)

  • Lee, Nam-Suk;Choi, Won-Suk;Qian, Dong-Jin;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2006.10a
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    • pp.91-92
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    • 2006
  • The electrical properties of viologen derivatives were studied in terms of the tunneling current characteristics on the length of the viologen derivatives using self-assembling techniques and ultra high vacuum scanning tunneling microscopy (UHV-STM). We fabricated the Au substrate were deposited by thermal evaporation system ($420^{\circ}C$. Self-assembled monolayers (SAMs) were prepared on Au (111), which had been thermally deposited onto freshly cleaved, heated mica. The Au substrate was exposed to a 1 mM solution of viologen derivatives in ethanol for 24 hours to form a monolayer. We measurement of the morphology on the single viologen molecules ($VC_{8}SH$, $VC_{10}SH$, $HSC_{8}VC_{8}SH$, and $HSC_{10}VC_{10}SH$). The current-voltage (I-V) and differential conductance (dl/dV-V) properties were measured while the electrical properties of the formed monolayer were scanned by using a STS. The effective barrier height of viologen derivatives ($VC_{8}SH$, $VC_{10}SH$, $HSC_{8}VC_{8}SH$, and $HSC_{10}VC_{10}SH$) were calculated to be 1.076 eV, 1.56 ${\pm}$ 0.3 eV, 1.85 eV, 2.28 eV, respectively.

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