• Title/Summary/Keyword: domain-specificity

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1H, 15N and 13C resonance assignment and secondary structure prediction of ss-DNA binding protein 12RNP2 precursor, HP0827 from Helicobacter pylori

  • Jang, Sun-Bok;Ma, Chao;Chandan, Pathak Chinar;Kim, Do-Hee;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.15 no.1
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    • pp.69-79
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    • 2011
  • HP0827 has two RNP motif which is a very common protein domain involved in recognition of a wide range of ssRNA/DNA.We acquired 3D NMR spectra of HP0827 which shows well dispersed and homogeneous signals which allows us to assign 98% of all $^1H_N$, $^{15}N$, $^{13}C_{\alpha}$, $^{13}C_{\beta}$ and $^{13}C$=O resonances and 90% of all sidechain resonances. The sequence-specific backbone resonance assignment of HP0827 can be used to gain deeper insights into the nucleic acids binding specificity of HP0827 in the future study. Here, we report secondary structure prediction of HP0827 derived from NMR data. Additionally, ssRNA/DNA binding assay studies was also conducted. This study might provide a clue for exact function of HP0827 based on structure and sequence.

Study on Preference Orientation and Effectiveness of an Integration Policy for the Rural married immigrant women according to life areas (생활영역별 농촌결혼이민여성 통합정책의 선호 정향성 및 효과성에 관한 연구)

  • Yang, Soon-Mi;Lee, Jin-Sook;Kim, Hai-Sook
    • Journal of Agricultural Extension & Community Development
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    • v.22 no.3
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    • pp.269-283
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    • 2015
  • This study aimed at identifying the effectiveness and preference orientation of an social integration policy for rural married immigrant women according to eight life areas. For this purpose was surveyed non-multicultural residents and multicultural couples. By analysis, frist, the assimilation orientation as an model of integration policy for the women was preferred most highly at the life language area by every respondent. The multiculturalism orientation was preferred most highly at the private area such as dietary-life culture. Second, ANOVA analysis showed that the positive effectiveness on the language policy are evaluated most highly by the respondent having assimilation preference orientation. And the positive effectiveness on dietary-life culture policy are evaluated most highly by the respondent having multiculturalism orientation. Third, positive effectiveness level of the policy estimated by the resident was lower than it of multicultural couple significantly. In conclusion, based on results of this study, it can be seen as on that point preference orientation of integration policy for the women, the domain specificity is present in the attitude of the multicultural couple and resident. Therefore it suspect that integration policy for the women should be considered differently according to life sphere.

Kindergarten and Elementary School Teachers′ Perception on the Gifted Children and the Education for the Gifted and talented (초등 및 유치원 교사의 영재교육에 대한 인식)

  • 박미영;이지현
    • Journal of Gifted/Talented Education
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    • v.11 no.3
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    • pp.155-173
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    • 2001
  • The purpose of this study was to investigate the kindergarten and elementary school teachers' perception on the gifted children and the education for the gifted and the talented. One hundred and fifteen kindergarten teachers and 116 elementary school teachers participated in this study as subjects. Teachers perceptions on the gifted children were generally right, except the relationships between the giftedness and birth order, the domain-specificity of the creativity, and the physical growth, sense of humor, and the socio-economic status of the gifted children. Teachers perceptions on the education for the gifted and talented also were generally right. Especially the elementary school teachers perceptions were higher than those of the kindergarten teachers.

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Rescuing p53 from mdm2 by a pre-structured motif in intrinsically unfolded SUMO specific protease 4

  • Kim, Do-Hyoung;Lee, Chewook;Kim, Bom;Lee, Si-Hyung;Han, Kyou-Hoon
    • BMB Reports
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    • v.50 no.10
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    • pp.485-486
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    • 2017
  • Many intrinsically unstructured/unfolded proteins (IUPs) contain transient local secondary structures even though they are "unstructured" in a tertiary sense. These local secondary structures are named "pre-structured motifs (PreSMos)" and in fact are the specificity determinants for IUP-target binding, i.e., the active sites in IUPs. Using high-resolution NMR we have delineated a PreSMo active site in the intrinsically unfolded mid-domain (residues 201-300) of SUMO-specific protease 4 (SUSP4). This 29-residue motif which we termed a p53 rescue motif can protect p53 from mdm2 quenching by binding to the p53-helix binding pocket in mdm2(3-109). Our work demonstrates that the PreSMo approach is quite effective in providing a structural rationale for interactions of p53-mdm2-SUSP4 and opens a novel avenue for designing mdm2-inhibiting anticancer compounds.

Expression and characterization of the recombinant human galectin-3 (유전자 재조합 Human galectin-3의 발현과 성상)

  • Kim, Byung-gyu;Woo, Hee-jong
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.547-554
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    • 1997
  • Galectin-3 is known as an animal ${\beta}$-galactoside-binding lectin charicterized with S-type carbohydrate recognition domain. It plays a role in growth, adherence and movement of cells. It is, also, related to the cell transformation and metastasis of tumor cells. In this study, we have expressed and purified recombinant human galectin-3 (rHgalectin-3) using E coli system and asialofetuin affinity chromatography for the future development of monoclonal antibody to Hgalectin-3, which is suggested as the tumor marker for the gastric and thyroid gland cancers. Expressed protein was confirmed as the Hgalectin-3 by immunoblot with cross-reactive murine monoclonal antibody. Lectin activity and specificity of purified protein were, also, confirmed by the competitive inhibition with galectin-3 specific carbohydrate, lactose. Like physiological galectin-3, lectin activity of the molecule was not changed in nonreduced condition. Dimer formation, furthermore, was observed at high concentration of the protein even in the reduced condition, which is well known in physiological galectin-3. These results showed purified rHgalectin-3 has the same activity and molecular nature compared to the physiological galectin-3.

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Signal Transduction of the Cytokine Receptor

  • Watanabe, Sumiko
    • Animal cells and systems
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    • v.2 no.2
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    • pp.153-164
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    • 1998
  • Cytokines regulate proliferation, differentiation and functions of haemotopoietic cells. Each cytokine possesses a variety of activities on various target cells (pleiotropy) and various cytokines have similar and overlapping activities on the same target cells (redundancy). The nature of these cytokine activities predicts unique feature of cytokine receptors, namely, cytokine has multiple receptors, different cytokines share a common receptor, and different cytokine receptors are linked to common signaling pathways. cDNA cloning of genes for cytokine receptors revealed distinct sets of receptor family with different structural features. The cytokine receptor superfamily consists of a largest family, and contains more than twenty cytokine receptor subunits. This receptor has common structural features in both extracellular and intracellular regions without tyrosine kinase domain. Another striking feature of the receptor is to share common subunit of multiple cytokines, which partly explains the redundancy of activities of some cytokines. Recent studies revealed detailed signaling events of the cytokine receptor, the primary activation of JAK and subsequent phosphorylation of tyrosine residues of receptor, and various cellular proteins. Many SH2 containing adapter proteins play an important role in cytokine signals, and this system has similarities with tyrosine kinase receptor signal transduction. STAT may mainly account for cytokine specific functions as suggested by knockout mice studies. It is of importance to note that cytokine activates multiple signaling pathways and the balance and combination of related signaling events may determine the specificity of functions of cytokines.

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Application of Engineered Zinc Finger Proteins Immobilized on Paramagnetic Beads for Multiplexed Detection of Pathogenic DNA

  • Shim, Jiyoung;Williams, Langley;Kim, Dohyun;Ko, Kisung;Kim, Moon-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1323-1329
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    • 2021
  • Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

Odorant Receptors Containing Conserved Amino Acid Sequences in Transmembrane Domain 7 Display Distinct Expression Patterns in Mammalian Tissues

  • Ryu, Sang Eun;Shim, Tammy;Yi, Ju-Yeon;Kim, So Yeun;Park, Sun Hwa;Kim, Sung Won;Ronnett, Gabriele V.;Moon, Cheil
    • Molecules and Cells
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    • v.40 no.12
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    • pp.954-965
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    • 2017
  • Mammalian genomes are well established, and highly conserved regions within odorant receptors that are unique from other G-protein coupled receptors have been identified. Numerous functional studies have focused on specific conserved amino acids motifs; however, not all conserved motifs have been sufficiently characterized. Here, we identified a highly conserved 18 amino acid sequence motif within transmembrane domain seven (CAS-TM7) which was identified by aligning odorant receptor sequences. Next, we investigated the expression pattern and distribution of this conserved amino acid motif among a broad range of odorant receptors. To examine the localization of odorant receptor proteins, we used a sequence-specific peptide antibody against CAS-TM7 which is specific to odorant receptors across species. The specificity of this peptide antibody in recognizing odorant receptors has been confirmed in a heterologous in vitro system and a rat-based in vivo system. The CAS-TM7 odorant receptors localized with distinct patterns at each region of the olfactory epithelium; septum, endoturbinate and ectoturbinate. To our great interests, we found that the CAS-TM7 odorant receptors are primarily localized to the dorsal region of the olfactory bulb, coinciding with olfactory epithelium-based patterns. Also, these odorant receptors were ectopically expressed in the various non-olfactory tissues in an evolutionary constrained manner between human and rats. This study has characterized the expression patterns of odorant receptors containing particular amino acid motif in transmembrane domain 7, and which led to an intriguing possibility that the conserved motif of odorant receptors can play critical roles in other physiological functions as well as olfaction.

The Effects of Field Trip Learning Program on Plant Inquiry in Coastal Dune using Artificial Intelligence on the Affective Domain of Gifted Elementary Science Studentt (인공지능을 활용한 해안사구 식물 탐구 프로그램이 초등 과학영재의 정의적 영역에 미치는 영향)

  • Byeon, Jung-Ho
    • Journal of Science Education
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    • v.46 no.1
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    • pp.53-65
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    • 2022
  • In the application and composition of learning content, the field trip learning of scientific inquiry could provide a positive effect. Also, it can arouse an experience of various inquiry activities through open thinking. In addition, it could take a positive effect by providing the diversity and specificity of wildlife experience for the living organism. The biology inquiry program of the field trip is a necessary process to acquire ecological experiences in the learning context. However, there is some problem to solve before the performance of field trip learning as professional knowledge of the outdoors inquiry. Therefore, this study developed a field trip inquiry program for the plant in a coastal dune using artificial intelligence to assist professional knowledge. The researcher carried out literature reviews and analysis related to studies and programs to investigate learning steps, content, and strategy. Also, this study investigated the effects of the program on the affective domain of gifted elementary science students. According to the results of this study, the program can provide a positive effect on motivation, task commitment, and attitude level. Consequently, the field trip learning program for plant in the coastal dune using artificial intelligence developed in this study can arouse a positive effect on the affective domain. Therefore, additional study is necessary related to inquiry programs of the field trip for various students and sites.

Kinetic Characterization of an Iron-sulfur Containing Enzyme, L-serine Dehydratase from Mycobacterium tuberculosis H37Rv (Mycobacterium tuberculosis H37Rv로부터 유래된 철-황 함유 효소인 L-세린 탈수화효소의 동력학적 특성)

  • Han, Yu Jeong;Lee, Ki Seog
    • Journal of Life Science
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    • v.28 no.3
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    • pp.351-356
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    • 2018
  • L-Serine dehydratase (LSD) is an iron-sulfur containing enzyme that catalyzes the conversion of L-serine to pyruvate and ammonia. Among the bacterial amino acid dehydratases, it appears that only the L-serine specific enzymes utilize an iron-sulfur cluster at their catalytic site. Moreover, bacterial LSDs are classified into four types based on structural characteristics and domain arrangement. To date, only the LSD enzymes from a few bacterial strains have been studied, but more detailed investigations are required to understand the catalytic mechanism of various bacterial LSDs. In this study, LSD type II from Mycobacterium tuberculosis (MtLSD) H37Rv was expressed and purified to elucidate the biochemical and catalytic properties using the enzyme kinetic method. The L-serine saturation curve of MtLSD exhibited a typically sigmoid character, indicating an allosteric cooperativity. The values of $K_m$ and $k_{cat}$ were estimated to be $59.35{\pm}1.23mM$ and $18.12{\pm}0.20s^{-1}$, respectively. Moreover, the plot of initial velocity versus D-serine concentration at fixed L-serine concentrations showed a non-linear hyperbola decay shape and exhibited a competitive inhibition for D-serine with an apparent $K_i$ value of $30.46{\pm}5.93mM$ and with no change in the $k_{cat}$ value. These results provide insightful biochemical information regarding the catalytic properties and the substrate specificity of MtLSD.