• Title/Summary/Keyword: 관 결합

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H2AX Directly Interacts with BRCA1 and BARD1 via its NLS and BRCT Domain Respectively in vitro (H2AX의 BRCA1 NLS domain과 BARD1 BRCT domain 각각과의 in vitro 상호 결합)

  • Bae, Seung-Hee;Lee, Sun-Mi;Kim, Su-Mi;Choe, Tae-Boo;Kim, Cha-Soon;Seong, Ki-Moon;Jin, Young-Woo;An, Sung-Kwan
    • KSBB Journal
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    • v.24 no.4
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    • pp.403-409
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    • 2009
  • H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.

Alternative Isoforms of the mi Transcription Factor (MITF) Regulate the Expression of mMCP-6 in the Connective Tissue-Type Mast Cells Cultured with Stem Cell Factor (SCF에서 배양한 결합조직형 비만세포에서 mMCP-6 발현을 조절하는 MITF 이형체)

  • Lee, Sun-Hee;Guan, Xiu-Ying;Kim, Dae-Ki
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1348-1354
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    • 2008
  • mi transcription factor (MITF) is important in regulating the differentiation of mast cells. In particular, MITF regulates the transcription of the mouse mast cell-specific serine protease (mMCP)-6 gene, which is generally expressed by the connective tissue-type of mast cells. In this study, we investigated alternative isoforms of MITF that regulate transcription of the mMCP-6 gene in bone marrow-derived cultured mast cells in mice. The expression of MITF isoforms was examined by RT-PCR. We observed that MITF-A, -E, -H and -Mc were expressed by mucosal-type mast cells cultured in the presence of IL-3, whereas the connective tissue-type mast cells cultured in the presence of stem cell factor (SCF) expressed MITF-A. Overexpression of MITF isoforms increased luciferase activity through the mMCP-6 promoter in NIH-3T3 cells and elevated the level of mMCP-6 expression in the MC/9 mast cell line. Moreover, mMCP-6 expression in mast cells was significantly inhibited by the depletion of MITF. The transcriptional activity and DNA binding of MITF-A was comparable to that of MITF isoforms, including MITF-E, -H, and -Mc. Our results therefore suggest that MITF-A may be an important isoform of MITF in regulating the transcription of mMCP-6 in mouse connective tissue mast cells.

Effect of Phosphate-to-binder and Water-to-binder Ratio on Magnesia-potassium Phosphate Cement (마그네시아-인산칼륨 시멘트에 대한 인산염 비 및 물-결합재비의 영향)

  • Lee, Kyung-Ho;Yoon, Hyun-Sub;Yang, Keun-Hyeok
    • Journal of the Korea Concrete Institute
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    • v.29 no.3
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    • pp.275-281
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    • 2017
  • This study examined the effect of water-to-binder ratio (W/B) and phosphate-to-binder ratio (P/B) on the flow, setting time, compressive strength development, and pH variation of magnesium-potassium phosphate composites, MKPC mortars. Ten mortars mixtures were prepared with the W/B varying from 20% to 40% at each P/B of 0.3 or 0.5. The hydration products and microstructural pore distribution of the MKPC pastes were investigated using X-ray diffraction (XRD), scanning electron microscope (SEM) and mercury intrusion porosimetry (MIP). The initial flow and setting time of MKPC mortars tended to decrease with an increase of P/B, indicating that the final setting time was shortened by approximately 24% when P/B increased from 0.3 to 0.5. The slope of the early-strength development measured in the MKPC mortars was considerably higher than that of cement concrete specified in code provisions. For obtaining a relatively good 28-day strength (above 30 MPa) and a near neutral pH (below 9.0) in MKPC mortars, the P/B and W/B need to be selected as 0.5 and 30%, respectively. The strubite-K crystal increased with the increases of P/B and W/B, which leads to the decrease of the macro-capillary pores.

Bandwidth Enhanced Miniaturization Method of Parallel Coupled-Line Filter (대역폭 특성이 개선된 평행 결합 선로 필터의 소형화 기법)

  • Myoung, Seong-Sik;Yook, Jong-Gwan
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.18 no.2 s.117
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    • pp.126-135
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    • 2007
  • This paper proposes a new miniaturization method for a parallel coupled line filter with enhanced bandwidth characteristics. A previous method incorporated several advantages, such as size reduction through the use of only a small number of capacitors, in addition to grounding, suppression of harmonic characteristics, and improved skirt characteristics for the parallel coupled line filter, which is conventional in the field of RE filters due to its design and fabrication simplicity. However, the previous method also has disadvantages related to the bandwidth shrinkage of the miniaturized filters. In this paper, the amount of bandwidth shrinkage is analyzed in terms of the relationship between the loaded Q(quality factor) and the group delay of a resonator. Moreover, the reduction in the bandwidth is solved by a design with new design equations. To show the validity of the proposed method, a hairpin filter with a center frequency of 5.2 GHz and an fractional bandwidth(FBW) of 10% was scaled down to half its original dimension by the proposed method with the enhanced bandwidth characteristics. The measured result shows a high level of agreement with theoretical results.

EFFECT OF CURING METHODS OF RESIN CEMENTS ON BOND STRENGTH AND ADHESIVE INTERFACE OF POST (레진시멘트의 중합방법이 포스트의 결합강도와 접착계면에 미치는 영향)

  • Kim, Mun-Hang;Kim, Hae-Jung;Cho, Young-Gon
    • Restorative Dentistry and Endodontics
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    • v.34 no.2
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    • pp.103-112
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    • 2009
  • The purpose of this study was to compare the effect of curing methods of adhesive resins and resin cements in the root canal. Crown portions of 32 single-rooted mandibular premolars were removed. Routine endodontic treatment was done, and 9 mm deep post spaces were prepared within root canals. No.3 FRC Postec posts (Ivoclar-Vivadent AG, Liechtensteih) were cemented in the post spaces by self-(SC) or light-curing (LC) using two dual-cured adhesives (Adper Scotchbond multi-purpose plus and Exite DSC )and resin cements (RelyX ARC and Variolink II). They were assigned to 4 groups (n=8): R-SC, R-LC, V-SC, V-LC group. After stored in distilled water for 24 hours, each root was transversally sectioned with 1.5 mm thick and made three slices. The specimens were subjected to push-out test in a universal testing machine (EZ Test, Shimadzu Co., Japan) with a crosshead speed of 1 mm/min. The data were analyzed with repeated ANOVA and one-way ANOVA. Also the interface of post-resin cement and resin cement-canal wall of each group was observed under FE-SEM. When fiber posts were cemented into the root canal using total-etch adhesives, the bond strength and adaptation between post and root canal dentin was affected by curing method. Self-cure of adhesives and resin cements showed higher bond strength and closer adaptation than light-cure of them.

Comparative Studies on the Ultrastructure of Salivary Ducts between the Two Species of Snails, Achatina fulica and Incilaria fruhstorferi (두 종의 달팽이류 (Achatina fulica and Incilaria fruhstorferi) 사이의 타액관의 미세구조에 관한 비교연구)

  • Chang, Nam-Sub;Han, Jong-Min;Kim, Sang-Won;Lee, Kwang-Ju;Hwang, Sun-Jong
    • Applied Microscopy
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    • v.30 no.1
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    • pp.89-100
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    • 2000
  • We observed the salivary ducts of two species of snails, Achatina fulica and Incilaria fruhstoferi with an electron microscope, and obtained the following results. The intralobular and interlobular ducts of Achatina fulica assume the forms of round or ellipsoidal doughnuts. The boundaries between the endothelial cells are not clear. It is also found that the cytoplasm of the endothelial cells consists of the membrane infolded in interdigital form, and there are well -developed microvilli at the apical portion of the cytoplasm. On the other hand, the intralobular and interlobular ducts of Incilaria fruhtoferi consist of the irregular simple columnar epithelia. The high electron dense cytoplasm is filled with the irregular round granules. The microvilli at the apical portion of the cytoplasm are not so well-developed as those in Achatina fulica. In the salivary duct of Achatina fulica, the lumen has narrow and long tubular structure. The boundaries between the endothelial cells are not clear. The cytoplasm is full of many vacuoles and electron lucent granules. At the apical portion of the cytoplasm, lots of short and thin microvilli are found. The salivary duct of Incilaria fruhstorferi is wider ($65\times250{\mu}m$ in diameter) than that of Achatina fulica, and consists of endothelial cells of the same structures. At the apical portion of those endothelial cells, a lot of junction apparatus such as desmosomes are observed. The vessels in the salivary ducts of Achatina fulica and Incilaria fruhstoferi are observed mainly in the connective tissues between the salivary glands. The endothelial cell of the vessel has the irregular structure and looks dark due to the high electron density. These cells protrude their filopodia and phagocytosize foreign bodies.

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Design of a Decentralized Controller for Deep-sea Mining System (심해저 채광시스템에 대한 분산제어기 설계에 관한 연구)

  • Yeu, Tae-Kyeong;Park, Soung-Jea;Hong, Sup;Kim, Hyung-Woo;Choi, Jong-Su
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.13 no.3
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    • pp.252-259
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    • 2008
  • The deep-sea mining system is generally composed of surface vessel, lifting system, buffer, flexible pipe and miner. The mining system can be regarded as a large-scale system in which each subsystem is interconnected to other ones. In order to control a large-scale system, decentralized control approaches have been proposed recently. In this paper, as a basic study on application of decentralized control, firstly, the mining system was modeled in a simplified way. Lifting system and buffer were regarded as a spherical pendulum and the flexible pipe was taken as a two-dimensional linear spring connection. Based on the simplified model dynamics, the mining system can be decentralized two subsystems, the one consisting of surface vessel, lifting system and buffer, and the other, the miner. Next, this paper proposed the design of controller for each decentralized subsystem by regarding the interacting terms as disturbances. The controllers kept the constant distance between two subsystems during the miner was moving on the specified track. Finally, the efficiency of proposed controller was proven through the numerical simulation of the derived model.

Effects of laser-irradiated dentin on shear bond strength of composite resin (레이저 처리가 상아질과 복합 레진의 결합에 미치는 영향)

  • Kim, Sung-Sook;Park, Jong-Il;Lee, Jae-In;Kim, Gye-Sun;Cho, Hye-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.5
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    • pp.520-527
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    • 2008
  • Purpose: This study was conducted to evaluate the shear bond strength of composite resin to dentin when etched with laser instead of phosphoric acid. Material and methods: Recently extracted forty molars, completely free of dental caries, were embedded into acrylic resin. After exposing dentin with diamond saw, teeth surface were polished with a series of SiC paper. The teeth were divided into four groups composed of 10 specimens each; 1) no surface treated group as a control 2) acid-etched with 35%-phosphoric acid 3) Er:YAG laser treated 4) Er,Cr:YSGG laser treated. A dentin bonding agent (Adapter Single Bond2, 3M/ESPE) was applied to the specimens and then transparent plastic tubes (3 mm of height and diameter) were placed on each dentin. The composite resin was inserted into the tubes and cured. All the specimens were stored in distilled water at $37^{\circ}C$ for 24 hours and the shear bond strength was measured using a universal testing machine (Z020, Zwick, Germany). The data of tensile bond strength were statistically analyzed by one-way ANOVA and Duncan's test at ${\alpha}$= 0.05. Results: The bond strengths of Er:YAG laser-treated group was $3.98{\pm}0.88$ MPa and Er,Cr:YSGG laser-treated group showed $3.70{\pm}1.55$ MPa. There were no significant differences between two laser groups. The control group showed the lowest bond strength, $1.52{\pm}0.42$ MPa and the highest shear bond strength was presented in acid-etched group, $7.10{\pm}1.86$ MPa (P < .05). Conclusion: Laser-etched group exhibited significantly higer bond strength than that of control group, while still weaker than that of the phosphoric acid-etched group.

Ferritin, an Iron Storage Protein, Associates with Kinesin 1 through the Cargo-binding Region of Kinesin Heavy Chains (KHCs) (철 저장 단백질 ferritin과 kinesin 1 결합 규명)

  • Jang, Won Hee;Jeong, Young Joo;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.26 no.6
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    • pp.698-704
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    • 2016
  • The intracellular transport of organelles and protein complexes is mediated by kinesin superfamily proteins (KIFs). The first kinesin, kinesin 1, was identified as a molecular motor protein that moves various organelles and protein complexes along the microtubule rails in cells. Kinesin 1 is a tetramer of two heavy chains (KHCs, also called KIF5s) and two kinesin light chains (KLCs). KIF5s interact with many different proteins through their tail region, but their binding proteins have not yet been fully identified. To identify the interaction proteins for KIF5A, we performed yeast two-hybrid screening and found a specific interaction with ferritin heavy chain (Frt-h), which has a role in iron storage and detoxification. Frt-h bound to the amino acid residues between 800 and 940 of KIF5A and to other KIF5s in the yeast two-hybrid assay. The coiled-coil domain of Frt-h is essential for interaction with KIF5A. In addition, ferritin light chain (Frt-l) interacted with KIF5s in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to KHC specifically co-immunoprecipitated Frt-h and Frt-l from mouse brain extracts. These results suggest the kinesin 1 motor protein may transport the ferritin complex in cells.

Glutamate-rich 4 Binds to Kinesin Superfamily Protein 5A (Glutamate-rich 4와 kinesin superfamily protein 5A와의 결합)

  • Se Young Pyo;Young Joo Jeong;Sung Woo Park;Mi Kyoung Seo;Won Hee Lee;Sang-Hwa Urm;Sang Jin Kim;Mooseong Kim;Jung Goo Lee;Dae-Hyun Seog
    • Journal of Life Science
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    • v.33 no.1
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    • pp.1-7
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    • 2023
  • Intracellular cargo transport is mediated by molecular motor proteins, such as kinesin and cytoplasmic dynein. Kinesins make up a large subfamily of molecular motors. Kinesin-1 is a plus-end-directed molecular motor protein that moves various cargoes, such as organelles, protein complexes, and mRNAs, along a microtubule track. It consists of the kinesin superfamily protein (KIF) 5A, 5B, and 5C (also called kinesin heavy chains) and kinesin light chains (KLCs). Kinesin-1 interacts with many different binding proteins through its carboxyl (C)-terminal region of KIF5s and KLCs, but their binding proteins have not yet been fully identified. In this study, a yeast two-hybrid assay was used to identify the proteins that interact with the KIF5A specific C-terminal region. The assay revealed an interaction between KIF5A and glutamate-rich 4 (ERICH4). ERICH4 bound to the KIF5A specific the C-terminal region but did not interact with the C-terminal region of KIF5B or KIF3A (a motor protein of kinesin-2). In addition, KIF5A did not interact with another isoform, ERICH1. Glutathione S-transferase (GST) pull-downs showed that KIF5A interacts with GST-ERICH4 and GST-ERICH4-amino (N)-terminal but not with GST-ERICH4-C or GST alone. When co-expressed in HEK-293T cells, ERICH4 co-localized with KIF5A and co-immunoprecipitated with KIF5A and KLC but not KIF3B. Together, our findings suggest that ERICH4 is capable of binding to KIF5A and that it may serve as an adaptor protein that links kinesin-1 with cargo.