• Title/Summary/Keyword: Domain interaction

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Genetic Screening of the Dazl-Interacting Protein Genes

  • Lee, Kyung-Ho;Lee, Seong-Ju;Rhee, Kun-Soo
    • Animal cells and systems
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    • v.10 no.4
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    • pp.227-231
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    • 2006
  • Micro-deletions at specific loci of the Y chromosome have been observed frequently in male infertility patients, suggesting that genes in these regions are involved in male germ cell development. DAZ is a representative male infertility gene at the AZFc locus of the Y chromosome. Since DAZ contains an RNA binding motif along with so-called a DAZ domain, it was proposed to participate in RNA metabolism during spermatogenesis. A mouse gene homologous to the human DAZ gene has been cloned and named Dazl (DAZlike). Dazl is autosomal and expressed in the testis and also at a low level in the ovary. Male mice homozygous for the Dazl null allele have small testes with a few spermatogonia and almost complete absence of germ cells beyond the spermatogonial stage, suggesting the requirement of Dazl for entry or progression through meiosis. However, its exact cellular functions have not been understood yet. In order to investigate cellular functions of Dazl, we decided to isolate candidate interacting protein genes of the mouse Dazl, using yeast two-hybrid screening. A number of candidate Dazlinteracting proteins have been isolated, such as Bprp, Acf, Hgs, Murr1, Nbak3 and Ranbp9, but dynein light chain 1 (Dlc1) was most predominant. A strong interaction of Dazl with Dlc1 suggests that Dazl might function as an mRNA adaptor to the dynein motor complex.

Identification of a Transferrin Receptor-binding Peptide from a Phage-displayed Peptide Library (파지-펩타이드 문고로부터 트랜스페린 수용체에 결합하는 펩타이드 탐색)

  • Kim, Sung-Il;Choi, Suk-Jung
    • Journal of Life Science
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    • v.18 no.3
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    • pp.298-303
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    • 2008
  • Using a phage peptide library approach, we have isolated a peptide ligand that binds to transferrin receptor on the surface of human melanoma cell, B16F10. The library was first screened twice by recovering internalized phages and was further screened three times by competitively eluting transferrin receptor-specific phages with human transferrin among the phages bound to the cell surface. The peptides displayed by the selected phages were fused to translocation and catalytic domain of Pseudomonas exotoxin to prepare recombinant toxins. After estimating cytotoxicity of each recombinant toxin toward B16F10 cell, seven clones were selected. Sequence analysis revealed that one of the clones displayed a peptide which had a significant sequence homology with human transferrin. The peptide was chemically synthesized and was shown to be functional in delivering cytotoxic agents into B16F10 cell via interaction with transferrin receptor.

Thermal Denaturation of the Apo-cyclic AMP Receptor Protein and Noncovalent Interactions between Its Domains

  • Won, Hyung-Sik;Seo, Min-Duk;Ko, Hyun-Suk;Choi, Wahn Soo;Lee, Bong-Jin
    • Molecules and Cells
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    • v.26 no.1
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    • pp.61-66
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    • 2008
  • Cyclic AMP receptor protein (CRP) is allosterically activated by cAMP and functions as a global transcription regulator in enteric bacteria. Structural information on CRP in the absence of cAMP (apo-CRP) is essential to fully understand its allosteric behavior. In this study we demonstrated interdomain interactions in apo-CRP, using a comparative thermodynamic approach to the intact protein and its isolated domains, which were prepared either by limited proteolysis or using recombinant DNA. Thermal denaturation of the intact apo-CRP, monitored by differential scanning calorimetry, revealed an apparently single cooperative transition with a slight asymmetry. Combined with circular dichroism and fluorescence analysis, the thermal denaturation of apo-CRP could be interpreted as a coupled process involving two individual transitions, each attributable to a structural domain. When isolated individually, both of the domains exhibited significantly altered thermal behavior, thus pointing to the existence of non-covalent interdomain interactions in the intact apo-CRP. These observations suggest that the allosteric conformational change of CRP upon binding to cAMP is achieved by perturbing or modifying pre-existing interdomain interactions. They also underline the effectiveness of a comparative approach using calorimetric and structural probes for studying the thermodynamics of a protein.

NMR structural studies on Human CD99 Type I

  • Kim, Hai-Young;Kim, Young-Mee;Joon Shin;Shin, Young-Kee;Park, Seong-Hoe;Lee, Weontae
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.69-69
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    • 2003
  • Human CD99 is a ubiquitous 32-kDa transmembrane protein encoded by the mic2 gene. The major cellular functions of CD99 protein are related to homotypic cell adhension, apoptosis, vesicular protein transport, and differentiation of thymocytes or T cells. Recently it has been reported that expression of a splice variant of CD99 transmembrane protein (Type I and Type II) increases invasive ability of human breast cancer cells. To understand structural basis for cellular functions of CD99 (Type I), we have initiated studies on hCD99$^{TMcytoI}$ and hCD99$^{cytoI}$ using circular dichroism (CD) and multi-dimensional NMR spectroscopy. CD spectrum of hCD99$^{TMcytoI}$ in the presence of 200mM DPC and CHAPS displayed an existence $\alpha$-helical conformation. The solution structure of hCD99$^{cytoI}$ determined by NMR is composed of one N-terminal $\alpha$-helix, $\alpha$A, two C-terminal short $\alpha$-helix segments, $\alpha$B and $\alpha$C. While $\alpha$A and $\alpha$B are connected by the long flexible loop, $\alpha$B and $\alpha$C connected by type III$\beta$-turn. Although it has been rarely figured out the correlation between structure and functional mechanism of hCD99$^{TMcytoI}$ and hCD99$^{cytoI}$, there is possibility of dimerization or oligomerization. In addition, the feasible mechanism of hCD99$^{cytoI}$ is that it could have intramolecular interaction between the N- and C- terminal domain through large flexible AB loop.

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Cobrotoxin Inhibits Prostate Carcinoma PC-3 Cell Growth Through Induction of Apoptotic Cell Death Via Inactivation of NF-kB

  • Song, Kyung-Chul;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.23 no.2
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    • pp.47-59
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    • 2006
  • We previously found that cobrotoxin inhibited $NF-{\kappa}B$ activity by reacting with signal molecules of $NF-{\kappa}B$ which is critical contributor in cancer cell growth by induction of apoptotic cell death. We here investigated whether cobrotoxin inhibits cell growth of human prostate cancer cells through induction of apoptotic cell death, which is related with the suppression of the $NF-{\kappa}B$ activity. Cobrotoxin $(0{\sim}8\;nM)$ inhibited prostate cancer cell growth through increased apoptosis in a dose dependent manner. Cobrotoxin inhibited DNA binding activity of $NF-{\kappa}B$, an anti-apoptotic transcriptional factor. Consistent with the induction of apoptosis and inhibition of $NF-{\kappa}B$, cobrotoxin increased the expression of pro-apoptotic proteins caspase 3. Cobrotoxin, a venom of Vipera lebetina turanica, is a group of basicpeptides composed of 233 amino acids with six disulfide bonds formed by twelve cysteins. NF-kB is activated by subsequent release of inhibitory IkB and translocation of p50. Since sulfhydryl group is present in kinase domain of p50 subunit of NF-kB, cobrotoxin could modify NF-kB activity by protein-protein interaction. And Cobrotoxin down regulated Akt signals. Salicylic acid as a reducing agent of Sulf-hydryl group and LY294002 as a Akt inhibitor abrogated cobrotoxin-induced cell growth and DNA binding activity of $NF-{\kappa}B$. These findings suggest that nano to pico molar range of cobrotoxin could inhibit prostate cancer cell growth, and the effect may be related with the induction of apoptotic cell death through Akt dependent inhibition of $NF-{\kappa}B$ signal.

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Effects of Education about Action Plans according to Self-Monitoring on Self-Management Adherence, Knowledge, Symptom Control, and Quality of Life among Adult Asthma Patients: A Randomized Controlled Trial (성인 천식환자의 자가감시 연계 활동계획서 교육이 자가관리 이행, 지식, 증상조절 및 삶의 질에 미치는 효과: 무작위대조시험설계)

  • Choi, Ja Yun;Kweon, Young-Ran
    • Journal of Korean Academy of Nursing
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    • v.47 no.5
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    • pp.613-623
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    • 2017
  • Purpose: This study was conducted to identify the effects of education regarding action plans according to a self-monitoring program on self-management adherence, knowledge, symptom control, and health-related quality of life (HRQoL) among adults with asthma. Methods: Thirty-four patients were randomly assigned to the intervention group and thirty-two to the control group in this study. A tailored 50-minute intervention based on the contents of self-monitoring and action plans developed by the National Heart Lung and Blood Institute was provided to the intervention group. Structured and well developed questionnaires were used to measure the dependent variables. Results: There were no differences in all general and clinical characteristics, and the dependent variables between two groups in the pre-test. In the post-test, there were differences in the level of self-management adherence (t=4.41, p<.001), knowledge (t=2.26, p=.027), symptom control (t=-2.56, p=.013), and total HRQoL (t=2.14, p=.036) between the two groups, although there was a difference only in the sub-domain of emotion (t=2.03, p=.047) in HRQoL. Conclusion: This study found that action plans according to self-monitoring that enhance a participatory interaction in the treatment and care could help patients with moderate to severe asthma to engagead equately in self-care, to control their symptoms, and to improve their HRQoL. Further studies are still needed to identify longitudinal effects of this program.

Case Study of Dynamic Amplification Characteristics of the Seismic Stations Using Observed Seismic Waves (관측지진파를 이용한 지반증폭특성 사례분석)

  • Lee, Jundae;Kim, Junkyoung
    • Journal of the Korean GEO-environmental Society
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    • v.10 no.1
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    • pp.35-41
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    • 2009
  • It is necessary to consider the site amplification for estimating SSI (soil structure interaction) and seismic source with more confidence. The horizontal to vertical (H/V) ratio technique in spectral domain is one of several techniques to estimate empirical site transfer function. The technique, originally proposed by Nakamura (1989), is applied to analyze the surface waves in the microtremor records. However, the application of this technique has been widened to the shear wave energy of strong motions for estimating site amplification. The purpose of this paper is to estimate spectral ratio using observed data at the seismic stations distributed within Southern Korean Peninsula from the Fukuoka earthquake including 11 aftershocks. The results show that each station has the its own characteristics of the specific resonance, high-band, and low-band frequency. The characteristics of the resonance frequency is more important because the quality of the seismic records are dependent on the resonance frequency. The result can be used for the study of site classification and removal of the site amplification effects from observed records can give us more reliable seismic source parameters.

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Regulation of signal transducer and activator of transcription 3 activation by dual-specificity phosphatase 3

  • Kim, Ba Reum;Ha, Jain;Kang, Eunjeong;Cho, Sayeon
    • BMB Reports
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    • v.53 no.6
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    • pp.335-340
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    • 2020
  • Since cancer is the leading cause of death worldwide, there is an urgent need to understand the mechanisms underlying cancer progression and the development of cancer inhibitors. Signal transducer and activator of transcription 3 (STAT3) is a major transcription factor that regulates the proliferation and survival of various cancer cells. Here, dual-specificity phosphatase 3 (DUSP3) was identified as a regulator of STAT3 based on an interaction screening performed using the protein tyrosine phosphatase library. DUSP3 interacted with the C-terminal domain of STAT3 and dephosphorylated p-Y705 of STAT3. In vitro dephosphorylation assay revealed that DUSP3 directly dephosphorylated p-STAT3. The suppressive effects of DUSP3 on STAT3 were evaluated by a decreased STAT3-specific promoter activity, which in turn reduced the expression of the downstream target genes of STAT3. In summary, DUSP3 downregulated the transcriptional activity of STAT3 via dephosphorylation at Y705 and also suppressed the migratory activity of cancer cells. This study demonstrated that DUSP3 inhibits interleukin 6 (IL-6)/STAT3 signaling and is expected to regulate cancer development. Novel functions of DUSP3 discovered in IL-6/STAT3 signaling regulation would help expand the understanding of cancer development mechanisms.

Numerical study of wake and aerodynamic forces on a twin-box bridge deck with different gap ratios

  • Shang, Jingmiao;Zhou, Qiang;Liao, Haili;Larsen, Allan;Wang, Jin;Li, Mingshui
    • Wind and Structures
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    • v.30 no.4
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    • pp.367-378
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    • 2020
  • Two-dimensional Delayed Detached Eddy Simulation (DDES) was carried out to investigate the uniform flow over a twin-box bridge deck (TBBD) with various gap ratios of L/C=5.1%, 12.8%, 25.6%, 38.5%, 73.3% and 108.2% (L: the gap-width between two girders, C: the chord length of a single girder) at Reynolds number, Re=4×104. The aerodynamic coefficients of the prototype deck with gap ratio of 73.3% obtained from the present simulation were compared with the previous experimental and numerical data for different attack angles to validate the present numerical method. Particular attention is devoted to the fluctuating pressure distribution and forces, shear layer reattachment position, wake velocity and flow pattern in order to understand the effects of gap ratio on dynamic flow interaction with the twin-box bridge deck. The flow structure is sensitive to the gap, thus a change in L/C thus leads to single-side shedding regime at L/C≤25.6%, and co-shedding regime at L/C≥35.8% distinguished by drastic changes in flow structure and vortex shedding. The gap-ratio-dependent Strouhal number gradually increases from 0.12 to 0.27, though the domain frequencies of vortices shedding from two girders are identical. The mean and fluctuating pressure distributions is significantly influenced by the flow pattern, and thus the fluctuating lift force on two girders increases or decreases with increasing of L/C in the single-side shedding and co-shedding regime, respectively. In addition, the flow mechanisms for the variation in aerodynamic performance with respect to gap ratios are discussed in detail.

Anti-IgE mAb Suppresses Systemic Anaphylaxis through the Inhibitory IgG Receptor Fc ${\gamma}$ RIIb in Mice - Interaction between Anti-IgE and Fc ${\gamma}$ RIIb -

  • Kang, Nam-In;Jin, Zhe-Wu;Lee, Hern-Ku
    • IMMUNE NETWORK
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    • v.7 no.3
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    • pp.141-148
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    • 2007
  • Background: Anti-IgE mAb which binds circulating but not receptor-bound IgE has been shown to be effective in treatment for asthma and other allergic diseases. However, the mechanisms by which anti-IgE mAb influences the pathophysiological responses are remained to be illustrated. This study was undertaken to examine the therapeutic efficacy of non-anaphylactogenic anti-mouse IgE mAb using murine models of IgE-induced systemic fatal anaphylaxis. Methods: Active systemic anaphylaxis was induced by either penicillin V(Pen V) or OVA and passive systemic anaphylaxis was induced by either anaphylactogenic anti-mouse IgE or a mixture of anti-chicken gamma globulin (CGG) IgG1 mAb and CGG. The binding of the Fc portion of anti-IgE to CHO-stable cell line expressing mouse Fc ${\gamma}$ RIIb was examined using flow cytometry. Fc fragments of anti-IgE mAb were prepared using papain digestion. The expression of phosphatases in lungs were assessed by Western blotting and immunohistochemistry. Results: Anti-IgE mAb prevented IgE- and IgG-induced active and passive systemic fatal reactions. In both types of anaphylaxis, anti-IgE mAb suppressed antigen-specific IgE responses, but not those of IgG. Anti-IgE mAb neither prevented anaphylaxis nor suppressed the IgE response in Fc ${\gamma}$ RIIb-deficient mice. The Fc portion of anti-IgE mAb was bound to murine Fc ${\gamma}$ RIIb gene-transfected CHO cells and inhibited systemic anaphylaxis. Anti-IgE mAb blocked the anaphylaxis-induced downregulation of Fc ${\gamma}$ RIIb-associated phosphatases such as src homology 2 domain-containing inositol 5-phosphatase (SHIP) and phosphatase and tensin homologue deleted on chromosome ten (PTEN). Conclusion: Anti-IgE mAb prevented anaphylaxis by delivering nonspecific inhibitory signals through the inhibitory IgG receptor, Fc ${\gamma}$ RIIb, rather than targeting IgE.