• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.45-45
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• 2003

We isolated a novel ankyrin-repeat containing protein, rSIAP (rSlo Interacting Ankyrin-repeat Protein), as an interacting protein to the cytosolic domain of the alpha-subunit of rat large-conductance Ca$\^$2+/-activated K$\^$+/ channel (rSlo) by yeast two-hybrid screening. Affinity pull-down assay showed the direct and specific interaction between rSIAP and rSlo domain. The channel-binding proteins can be classified into several categories according to their functional effects on the channel proteins, i.e. signaling adaptors, scaffolding net, molecular tuners, molecular chaperones, etc. To obtain initial clues on its functional roles, we investigated the cellular localization of rSIAP using immunofluorescent staining. The results showed the possible co-localization of rSlo and rSIAP protein near the plasma membrane, when co-expressed in CHO cells. We then investigated the functional effects of rSIAP on the rSlo channel using electrophysiological means. The co-expression of rSIAP accelerated the activation of rSlo channel. These effects were initiated at the micromolar [Ca$\^$2+/]$\_$i/ and gradually increased as [Ca$\^$2+/]$\_$i/ raised. Interestingly, rSIAP decreased the inactivation kinetics of rSlo channel at micromolar [Ca$\^$2+/]$\_$i/, while the rate was accelerated at sub-micromolar [Ca$\^$2+/]$\_$i/. These results suggest that rSIAP may modulate the activity of native BK$\_$Ca/ channel by altering its gating kinetics depending on [Ca$\^$2+/]$\_$i/. To localize critical regions involved in protein-protein interaction between rSlo and rSIAP, a series of sub-domain constructs were generated. We are currently investigating sub-domain interaction using both of yeast two-hybrid method and in vitro binding assay.

• 한국자기공명학회논문지
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• 제13권1호
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• pp.56-63
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• 2009

SWIRM domain, a core domain of SRG3 is well conserved in SW13, RSC8, and MOIRA family proteins. To understand structural basis for cellular functions of the SWIRM domain, we have initiated biochemical and structural studies on SWIRM domain and mutants using gelfiltration chromatography, circular dichroism and NMR spectroscopy. The structural properties of the mutant SWIRM domains (K34A and M75A) have been characterized, showing that the structures of both wild-type and mutant proteins are a-helical conformation. The data conclude that mutations at interaction sites of its binding partner protein do not affect its secondary and tertiary structure.

• 제어로봇시스템학회:학술대회논문집
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• 제어로봇시스템학회 2005년도 ICCAS
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• pp.107-112
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• 2005

Recently proposed stable teleoperation control scheme, base on time domain passivity, is modified to remove several conservatisms. During unconstrained motion and contacting with soft and deformable environments, the two-port time domain passivity approach [21] was excessively dissipating energy even though it was stable without any energy dissipation. The main reason of this conservatism is on the fact that the time domain passivity controller does not include the external energy dissipation elements at the slave manipulator. The measured interaction force between slave and environment allow the time domain passivity observer to include the amount of energy dissipation of the slave manipulator to the monitored energy. With the modified passivity observer, reference energy following idea [24] is applied to satisfy the passivity condition. The feasibility of the developed methods is proved with experiments. Improved performance is obtained in unconstrained motion and contacting with a soft environment.

• Interaction and multiscale mechanics
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• 제2권1호
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• pp.91-107
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• 2009

In order to accurately estimate the seismic behavior of buildings, it is important to consider both nonlinear characteristics of the buildings and the frequency dependency of the soil impedance. Therefore, transform methods of the soil impedance in the frequency domain to the impulse response in the time domain are needed because the nonlinear analysis can not be carried out in the frequency domain. The author has proposed practical transform methods. In this paper, seismic response analyses considering frequency dependent soil impedance in the time domain are shown. First, the formulation of the proposed transform methods is described. Then, the linear and nonlinear earthquake response analyses of a building on 2-layered soil were carried out using the transformed impulse responses. Through these analyses, the validity and efficiency of the methods were confirmed.

• 한국철도학회:학술대회논문집
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• 한국철도학회 2004년도 추계학술대회 논문집
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• pp.800-805
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• 2004

In this paper, a numerical method for train/track/structure interaction analysis in frequency domain is developed. Track is modelled as continuous beams supported by elastomers. The motion of train is expressed by those of car body, bogies and unsprung masses supported by springs and dampers. The equation of motion for train and track interaction system is derived by applying compatibility condition at the contact points between wheels and rails. The Effects of rail pad stiffness on the behaviors of train and track are analyzed using the presented method.

• 한국지진공학회:학술대회논문집
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• 한국지진공학회 2003년도 춘계 학술발표회논문집
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• pp.415-421
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• 2003

With the increasing possibility of earthquake occurrence, seismic safety of bridges has become one of the most important social issues in Korea. In this study, a nonlinear earthquake response analysis is carried out for a real bridge by incorporating soil-structure interaction and pier nonlinearity. The material nonlinearity of the bridge pier is realized by utilizing SAP2000 whereas the soil-structure interaction is analized in time domain by adapting KIESSI. The numerical results are compared to those of the models without considering the effects.

• 융합정보논문지
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• 제8권1호
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• pp.115-121
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• 2018

융복합시대에 유치원교사의 권리와 의무가 교사-유아 상호작용에 미치는 영향을 살펴보는 것이 본 연구의 목적이다. 서울, 경기, 인천 지역의 유치원 재직중인 교사 174명을 임의표집하여 설문하고 연구분석을 실시하였다. 조사결과, 교사의 권리가 교수-유아 상호작용의 하위영역인 언어적 상호작용에서는 교육과 신분상의 권리는 정적인 영향을 미쳤고, 정서적 상호작용에서는 교육권리에서만 정적인 영향을 미쳤다. 그리고 행동적 상호작용에서는 노동의권리가 부적인 영향을 미쳤다. 교사의 의무가 교수-유아 상호작용의 하위영역인 언어적 상호작용과 정서적 상호작용에서는 신분상의 의무는 정적인 영향을 미쳤고, 교사의 의무가 교수-유아 상호작용의 하위영역인 행동적 상호작용에서는 성실 및 직무상의 의무와 신분상의 의무는 영향을 미치지 않았다.

• Molecules and Cells
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• 제41권9호
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• pp.842-852
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• 2018

Notch signaling is an evolutionarily conserved pathway and involves in the regulation of various cellular and developmental processes. Ligand binding releases the intracellular domain of Notch receptor (NICD), which interacts with DNA-bound CSL [CBF1/Su(H)/Lag-1] to activate transcription of target genes. In the absence of NICD binding, CSL down-regulates target gene expression through the recruitment of various corepressor proteins including SMRT/NCoR (silencing mediator of retinoid and thyroid receptors/nuclear receptor corepressor), SHARP (SMRT/HDAC1-associated repressor protein), and KyoT2. Structural and functional studies revealed the molecular basis of these interactions, in which NICD coactivator and corepressor proteins competitively bind to ${\beta}-trefoil$ domain (BTD) of CSL using a conserved ${\varphi}W{\varphi}P$ motif (${\varphi}$ denotes any hydrophobic residues). To date, there are conflicting ideas regarding the molecular mechanism of SMRT-mediated repression of CSL as to whether CSL-SMRT interaction is direct or indirect (via the bridge factor SHARP). To solve this issue, we mapped the CSL-binding region of SMRT and employed a 'one- plus two-hybrid system' to obtain CSL interaction-defective mutants for this region. We identified the CSL-interaction module of SMRT (CIMS; amino acid 1816-1846) as the molecular determinant of its direct interaction with CSL. Notably, CIMS contains a canonical ${\varphi}W{\varphi}P$ sequence (APIWRP, amino acids 1832-1837) and directly interacts with CSL-BTD in a mode similar to other BTD-binding corepressors. Finally, we showed that CSL-interaction motif, rather than SHARP-interaction motif, of SMRT is involved in transcriptional repression of NICD in a cell-based assay. These results strongly suggest that SMRT participates in CSL-mediated repression via direct binding to CSL.

• Archives of Pharmacal Research
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• 제26권10호
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• pp.846-854
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• 2003

The oligomerization of G-proteincoupled receptors (GPCRs) has been shown to occur by various mechanisms, such as via disulfide covalent linkages, non covalent (ionic, hydrophobic) interactions of the N-terminal, and/or transmembrane and/or intracellular domains. Interactions between GPCRs could involve an association between identical proteins (homomers) or non-identical proteins (heteromers), or between two monomers (to form dimers) or multiple monomers (to form oligomers). It is believed that muscarinic receptors may also be arranged into dimeric or oigomeric complexes, but no systematic experimental evidence exists concerning the direct physical interaction between receptor proteins as its mechanism. We undertook this study to determine whether muscarinic receptors form homomers or a heteromers by direct protein-protein interaction within the same or within different subtypes using a yeast two-hybrid system. Intracellular loops (i1, i2 and i3) and the C-terminal cytoplasmic tails (C) of human muscarinic (Hm) receptor subtypes, Hm1, Hm2 and Hm3, were cloned into the vectors (pB42AD and pLexA) of a two-hybrid system and examined for heteromeric or homodimeric interactions between the cytoplasmic domains. No physical interaction was observed between the intracellular domains of any of the Hm/Hm receptor sets tested. The results of our study suggest that the Hm1, Hm2 and Hm3 receptors do not form dimers or oligomers by interacting directly through either the hydrophilic intracellular domains or the C-terminal tail domains. To further investigate extracellular domain interactions, the N-terminus (N) and extracellular loops (o1 and o2) were also cloned into the two-hybrid vectors. Interactions of Hm2N with Hm2N, Hm2o1, Hm2o2, Hm3N, Hm3o1 or Hm3o2 were examined. The N-terminal domain of Hm2 was found to have no direct interaction with any extracellular domain. From our results, we excluded the possibility of a direct interaction between the muscarinic receptor subtypes (Hm1, Hm2 and Hm3) as a mechanism for homo- or hetero-meric dimerization/oligomerization. On the other hand, it remains a possibility that interaction may occur indirectly or require proper conformation or subunit formation or hydrophobic region involvement.

• BMB Reports
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• 제31권4호
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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.