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Grim Stimulates Diap1 Poly-Ubiquitination by Binding to UbcD1

  • Yoo, Soon Ji
    • Molecules and Cells
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    • v.20 no.3
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    • pp.446-451
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    • 2005
  • Diap1 is an essential Drosophila cell death regulator that binds to caspases and inhibits their activity. Reaper, Grim and Hid each antagonize Diap1 by binding to its BIR domain, activating the caspases and eventually causing cell death. Reaper and Hid induce cell death in a Ring-dependent manner by stimulating Diap1 auto-ubiquitination and degradation. It was not clear that how Grim causes the ubiquitination and degradation of Diap1 in Grim-dependent cell death. We found that Grim stimulates poly-ubiquitination of Diap1 in the presence of UbcD1 and that it binds to UbcD1 in a GST pull-down assay, so presumably promoting Diap1 degradation. The possibility that dBruce is another E2 interacting with Diap1 was examined. The UBC domain of dBruce slightly stimulated poly-ubiquitination of Diap1 in Drosophila extracts but not in the reconstitution assay. However Grim did not stimulate Diap1 poly-ubiquitination in the presence of the UBC domain of dBruce. Taken together, these results suggest that Grim stimulates the poly-ubiquitination and presumably degradation of Diap1 in a novel way by binding to UbcD1 but not to the UBC domain of dBruce as an E2.

Identification of binding motifs for skeletal ryanodine receptor and triadin

  • Lee, Jae-Man;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.66-66
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    • 2003
  • In skeletal muscle cells, depolarization of the transverse tubules (T-tubules) results in Ca$\^$2+/ release from the sarcoplasmic reticulum (SR), leading to elevated cytoplasmic Ca$\^$2+/ and muscle contraction. This process has been known as excitation-contraction coupling (E-C coupling). Several proteins, such as the ryanodine receptor (RyR), triadin, junctin, and calsequestrin (CSQ), have been identified to be involved in the Ca$\^$2+/ release process. However, the molecular interactions between the SR proteins have not been resolved. In the present study, the mechanisms of interaction between RyRl and triadin have been studied by in vitro protein binding and $\^$45/Ca$\^$2+/ overlay assays. Our data demonstrate that the intraluminal loop II of RyR1 binds to triadin in Ca$\^$2+/-independent manner. Moreover, we could not find any Ca$\^$2+/ binding sites in the loop II region. GST-pull down assay revealed that a KEKE motif of triadin, which was previously identified as a CSQ binding site (Kobayasi et al.,2000 JBC) was also a binding site for RyR1. Our results suggest that the intraluminal loop II of RyR could participate in the RyR-mediated Ca$\^$2+/ release process by offering a direct binding site to luminal triadin.

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Molecular Cloning and Characterization of Calumenin in Rabbit Skeletal Sarcoplasmic Reticulum

  • Jung, Dai-Hyun;Kim, Do-Han
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.53-53
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    • 2003
  • Calumenin was previously identified as a high affinity Ca$\^$2+/ binding protein in mouse cardiac sarcoplasmic reticulum (SR). For the present study, a 48 kDa skeletal homologue of calumenin was identified by sucrose-density gradient of rabbit skeletal SR membranes, concanavalin A treatment, 2D-gel electrophoresis, $\^$45/Ca$\^$2+/ overlay, Stains-all staining, and MALDI-TOF analysis. We attempted to clone the skeletal calumenin by RT-PCR based on mouse cardiac and human calumenin sequences. The deduced amino acid sequence (315 residues) of the skeletal calumenin showed high identity to mouse cardiac calumenin (90%). As seen in the cardiac calumenin, the deduced sequence contains a 19 amino acid N-terminal signal sequence and a HDEF C-terminal sequence, a putative retrieval signal to ER. Also, the skeletal calumenin contains one N-glycosylation site, three PKC phosphorylation sites, eight casein kinase 2 phosphorylation sites, and 6 EF-hand domains. GST-calumenin showed a conformational change and increased mobility in the presence of Ca$\^$2+/ in SDS-PAGE. Three calumenin interacting proteins (ryanodine receptor 1, glycogen phosphorylase, and phosphofructo kinase) were identified by pull-down assay with GST-calumenin and solubilized SR. All the interactions were Ca$\^$2+/dependent. The present results suggest that calumenin plays an important role in Ca$\^$2+/ homeostasis of muscle cells.

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Ankyrin-B Interacts with the C-terminal Region of Hsp40

  • Min, Byung-In;Ko, Han-Suk;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.9 no.2
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    • pp.105-110
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    • 2003
  • Ankyrins are a ubiquitously expressed family of intracellular adaptor proteins involved in targeting diverse proteins to specialized membrane domains in both the plasma membrane and the endoplasmic reticulum. Canonical ankyrins are 190-220 kDa proteins expressed in most tissues and cell types and comprise a membrane-binding domain (MBD) of 24 ANK repeats, a spectrin-binding domain, a death domain and a C-terminal domain. Rescue studies with ankyrin-B/G chimeras have identified the C-terminal domain of ankyrin-B as the defining domain in specifying ankyrin-B activity, but the function of C-terminal domain of ankyrin-B is, however, not known. We report here that the C-terminal domain of ankyrin-B is capable of interacting with the C-terminal Region of Hsp40. The Hsps are induced not only by heat shock but also by various other environmental stresses. Hsps are also expressed constitutively at normal growth temperatures and have basic and indispensable functions in the life cycle of proteins as molecular chaperones, as well as playing a role in protecting cells from the deleterious stresses. The binding sites required in the interaction between C-terminal domain of ankyrin-B and C-terminal region of Hsp40 were characterized using the yeast two-hybrid system and GST-pull down assay. The interaction between ankyrin-B and Hsp40 represents the first direct evidence of ankyrin's role as chaperones.

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Identification and characterization of a rice MCM2 homologue required for DNA replycation

  • Cho, Jae-Han;Kim, Ho-Bang;Kim, Hyung-Sae;Choi, Sang-Bong
    • BMB Reports
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    • v.41 no.8
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    • pp.581-586
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    • 2008
  • The pre-replication complex (pre-RC), including the core hexameric MCM2-7 complex, ensures that the eukaryotic genome is replicated only once per cell division cycle. In this study, we identified a rice $\underline{m}ini\underline{c}hromosome$ $\underline{m}aintenance$ (MCM) homologue (OsMCM2) that functionally complemented fission yeast MCM2 (CDC19) mutants. We found OsMCM2 transcript expression in roots, leaves, and seeds, although expression levels differed slightly among the organs. Likewise, the OsMCM2 protein was ubiquitously expressed, but it was downregulated when nutritients were limiting, indicating that MCM2 expression (and therefore cell cycle progression) requires adequate nutrition. Yeast two-hybrid and GST pull-down assays demonstrated that OsMCM2 interacted with the COP9 signalosome 5 (CSN5). Taken as a whole, our results indicated that OsMCM2 functions as a subunit of the rice MCM complex and interacts with CSN5 during developmental regulation.

Development of a Hydraulic Servo Cylinder with an Integrated Feedback Mechamism (일체형 파드백 기구를 갖는 유압 서보실린더 개발 연구)

  • Lee, Jae-Gyu;Kim, Ock-Hyun
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.20 no.8
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    • pp.2480-2490
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    • 1996
  • This paper presents a new type of hydraulic servo chllinder which is characterized by its simple construction and an ubtegrated feedback mechanism. Piston position of the cylinder is controlled by eletrical input and mechamical feedback deduced from its own structure. Hydraulic pressure in each cylinder room is controlled by a poppet valve. The poppet is activated by a solenoid and is linked to the piston. Solenoid input current pulls up the poppet, which results in pressure drop and thus piston motion. The piston motion generates pull down force on the poppet by the linkage and the motion stops at equilibrium. In that way the piston position is controlled by an expernal input current. Characteristics of the servo cylinder is verified by stability analysis, tranient vehavior and steady state positing for step input. Design parameter analyses have been executed by derivation of analytical approximate solutions and by computer simulations. A prototype hydraulic servo cylinder is developed and tested. The experimental results show successful function of the servo cylinder and consistency with the theoritical results.

Interaction between Information Structure and Menu Design on Information Searching and Attitude in WWW (인터넷 정보탐색 과정에서 정보구조와 메뉴디자인의 상호작용 분석)

  • Yu Byeong-Min
    • The KIPS Transactions:PartB
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    • v.12B no.4 s.100
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    • pp.473-478
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    • 2005
  • The purpose of this study was to investigate the interaction effect of Web site menu designs and information structures on two information searching performances (searching and browsing) and three measures of users' attitudes (appeal, usability, and disorientation). Pronounced interaction effects occurred across four dependent variables except searching when decreasing and increasing information structure were combined with a simple selection menu and a pull-down menu. Further studies are needed to investigate additional interactions among factors of interface and information structure of Web sites.

The Optimized Monolithic Fault Protection Circuit for the Soft-shutdown behavior of 600V PT-IGBT by employing a New Blanking Filter (600V Punch-through형 절연 게이트 바이폴라 트랜지스터의 Soft-shutdown을 위해 시간 지연 회로를 적용한 새로운 보호회로)

  • Lim, Ji-Yong;Ji, In-Hwan;Ha, Min-Woo;Choi, Young-Hwan;Choi, Yearn-Ik;Han, Min-Koo
    • Proceedings of the KIEE Conference
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    • 2006.07c
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    • pp.1299-1300
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    • 2006
  • Floating p-well 전압 감지를 이용한 시간 지연 회로를 적용하여 punch-though형 절연 게이트 바이폴라 트랜지스터 (PT-IGBT)의 최적화된 보호 회로를 제안하였다. Floating P-well 축전기와 게이트 저항은 정상 스위칭 동작시 단락 회로 감지 동작 (false detection of fault)을 차단하며, floating p-well 전압은 단락회로 상황시 풀-다운 (pull-down) MOSFET의 문턱 전압 이상으로 상승되어 풀-다운 MOSFET을 턴-온(turn-on) 시킴으로서 IGBT의 게이트 전압을 감소시킨다. 이에 따라 IGBT의 컬렉터 전류는 자연스럽게 감소된다. 실험 결과를 통해, 단락회로 상황에서 최적화된 IGBT의 보호회로가 소프트-셧다운 (soft-shutdown) 특성을 보이는 것을 확인할 수 있다.

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The design and implementation of automatic translation system for hangul's romanization (국어 로마자 표기 자동 변환 시스템 설계 및 구현)

  • 김홍섭
    • KSCI Review
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    • v.2 no.1
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    • pp.45-54
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    • 1995
  • This study is, by assigning ASCII codes hardly used to Bandaljum(ˇ) and making the fonts of Korean-English character mode, to design the way of converting automatically a word, a sentence or a document of korean into phonetic letters by applying the algorismized phonological principles inputted as a letter string, even though a user do not konw the basic principles of the usage of Korean-to-Romanization notation rule. This is designed so that it may be possible to turn into a mechanical code with reference to the corresponding character in the table of Korean-to-Romanization notation rule that is the currently used standard proposition of the government. Consequently this program makes it user more convenient in the manipulations of special case words, the assistance of colorful-screen or pull-down, pop-up menu and the adoptation of utilizable mouse works for a user convienency. This program could be installed in a single diskette of 5.25"(2HD) and be made in C programming language to mplement various font, expansion or condense of font, alternative printing.ting.

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The residual mechanical properties evaluation according to temperature of the amorphous metallic fiber reinforced high strength concrete (비정질강섬유 보강 고강도 콘크리트의 온도별 잔존 역학적 특성 평가)

  • Suh, Dong-Kyun;Kim, Gyu-Yong;Lee, Sang-Kyu;Hwang, Eui-Chul;Eu, Ha-Min;Nam, Jeong-Soo
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2020.11a
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    • pp.98-99
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    • 2020
  • This study is aim to assess mechanical properties which is highly related to structural safe and durability of 100MPa high strength concrete mixed with amorphous metallic fiber. All specimens were heated with low velocity heating rate(1℃/min.), residual compressive strength and residual flexural strength was evaluated. The specimens were cooled down to room temperature after heating. As a result, in the case of 100MPa high-strength concrete, the residual compressive strength enhancing effect of amorphous metallic fiber has showed with the mix proportion of fiber. In addition, residual flexural strength showed more regular pattern before 300℃ then residual compressive strength, but simillar decreasing behavior was shown after 300℃ like residual compressive strength. Further study about fiber pull-out behavior and fiber mechanical, chemical property change due to temperature is needed.

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