• Archives of Pharmacal Research
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• 제20권5호
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• pp.454-458
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• 1997

Two human liver UDP-glucuronosyltransferase cDNA clones, HLUG25 and UDPGTh2 were previously shown to encode isozymes active in the glucuronidation of hyodeoxycholic acid (HDCA) and certain estrogen derivatives (e.g., estriol and 3,4-catechol estrogens), respectively. in this study we have found that the UDPGTh2-encoded isoform (UDPGTh2) and HLUG25-encoded isoform (UDPGThl) have parallel aglycone specificities. When expressed in COS 1 cells, each isoform metabolized three types of dihydroxy- or trihydroxy-substituted ring structures, including the 3,4-catechol estrogen (4-hydroxyestrone), estriol, 17-epiestriol, and HDCA, but the UDPGTh2 isozyme was 100-fold more efficient than UDPGTh1. UDPGTh1 and UDPGTh2 were 86% identical overall (76 differences out of 528 amino acids), including 55 differences in the first 300 amino acids of the amino terminus, a domain which conferred the substrate specificity. The data indicated that a high level of conservation in the amino terminus was not required for the preservation of substrate selectivity. Analysis of glucuronidation activity encoded by UDPGTh1/UDPGTh2 chimeric cDNA constructed at their common restriction sites, Sac I (codon 297), Nco I (codon 385), and Hha I (codon 469), showed that nine amino acids between residues 385 and 469 were important for catalytic efficiency, suggesting that this region represented a domain which was critical for the catalysis but distinct from that responsible for aglycone-selection. These data indicate that UDPGTh2 is a primary isoform responsible for the detoxification of the bile salt intermediate as well as the active estrogen intermediates.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6685-6689
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• 2014

Background: Alterations in gene expression levels or mutations of tyrosine kinases are detected in some human cancers. In this study, we examined whether serine threonine tyrosine kinase 1 (STYK1)/novel oncogene with kinase domain (NOK) is overexpressed in patients with colorectal cancer. We also examined the clinical relevance of STYK1/NOK expression in cancer tissues. Materials and Methods: In tumor samples of patients with colorectal cancer and their matched non-cancerous samples, STYK1/NOK messenger RNA (mRNA) expression was analyzed by quantitative reverse transcriptase polymerase chain reaction. Associations between the expression levels of STYK1/NOK and clinicopathological characteristics of colorectal cancer were also assessed using Mann-Whitney U and Kruskal-Wallis tests. Results: Upregulation of STYK1/NOK was found in cancer tissues even at early stage of colorectal cancer compared to normal adjacent tissues. The optimal cutoff point of 0.198 the STYK1/NOK expression showed 0.78 sensitivity and 0.75 specificity for diagnosis. Overexpressed STYK1/NOK was correlated with tumor size but had no association with other clinicopathological characteristics of colorectal cancer. Conclusions: These results indicate that STYK1/NOK mRNA is widely expressed in the patients with colorectal cancer and suggest that inhibition of this molecule could potentially serve as a novel therapeutic target.

• 컴퓨터교육학회논문지
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• 제16권6호
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• pp.11-19
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• 2013

최근 창의성의 영역 보편성 및 영역 특수성에 대한 논의가 계속되어지고 있는 시점에서 컴퓨터 프로그래밍 학습 활동 또한 창의성과 관련하여 다시 연구되어질 필요가 있다. 기존 교육용 프로그래밍 언어학습의 창의성에 대한 측정을 TTCT 도형 검사를 사용한 것에 반해, 본 연구에서는 프로그래밍 활동이 언어적 사고 양식을 더욱 요구하는 창의적 활동이라는 관점으로 바라보고 있다. 이에 따라, 본 연구에서 컴퓨터 과학의 핵심 개념을 학습 내용으로 하는 창의적 문제 해결(CPS) 모형 기반의 스크래치 프로그래밍 학습을 개발하였다. 이 학습 프로그램을 4, 5학년 학생 17명을 대상으로 5일간 4차시씩 총 20차시 적용하고, 정규성이 검증된 이 실험집단에 TTCT 언어 검사의 평균 표준점수(100)로 t검정을 실시한 결과 언어 창의성의 세가지 하위 요인(유창성, 융통성, 독창성)과 창의성 지수(세 요인의 평균 표준점수)가 유의미하게 증진되었다.

• BMB Reports
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• 제41권6호
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• pp.415-434
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• 2008

Phosphoinositide-specific phospholipase C is an effector molecule in the signal transduction process. It generates two second messengers, inositol-1,4,5-trisphosphate and diacylglycerol from phosphatidylinositol 4,5-bisphosphate. Currently, thirteen mammal PLC isozymes have been identified, and they are divided into six groups: PLC-$\beta$, -$\gamma$, -$\delta$, -$\varepsilon$, -$\zeta$ and -$\eta$. Sequence analysis studies demonstrated that each isozyme has more than one alternative splicing variant. PLC isozymes contain the X and Y domains that are responsible for catalytic activity. Several other domains including the PH domain, the C2 domain and EF hand motifs are involved in various biological functions of PLC isozymes as signaling proteins. The distribution of PLC isozymes is tissue and organ specific. Recent studies on isolated cells and knockout mice depleted of PLC isozymes have revealed their distinct phenotypes. Given the specificity in distribution and cellular localization, it is clear that each PLC isozyme bears a unique function in the modulation of physiological responses. In this review, we discuss the structural organization, enzymatic properties and molecular diversity of PLC splicing variants and study functional and physiological roles of each isozyme.

• KSBB Journal
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• 제19권2호
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• pp.159-163
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• 2004

$\AA$의 분해능을 가진 Bacillus macerans에서 분리 정제한 cyclodextrin glucanotransferase의 결정 구조를 X-ray를 이용하여 분자 대치법을 사용하여 밝혔다. 정확한 구조는 16.6% ( $R_{free}$ = 20.5%)의 결정학적 R-factor를 가지고 있다. 두 개의 $Ca^{2+}$$_{-}$/ 이온이 점유된 새로운 금속 결합자리는 활성 자리의 도달 채널에서 발견되었다. $Ca^{2+}$$_{-}$/ 이온에 결합하는 음의 하전을 띤 아미노산 잔기들이 많이 밀집되어 있고 이 때, domain A ($\alpha$H에서 아미노산 잔기 283-297)의 중심부에 연결 부분은 $\beta$ 13-$\alpha$G 이었다. $\beta$13-$\alpha$G 부분은 활성 자리의 입구에 위치한 subsite 1에서 -1 과 Tyr260 (subsite 2)사이에 있는 촉매 부위 Glu258을 포함하고 있다. 비록 $\alpha$-CGTase 부류에서 잘 보존되어 있지는 않으나, 새로운 $Ca^{2+}$$_{-}$/ 자리 3a,b가 이 $\alpha$-CGTase의 활성도와 생성물의 특이성에 중요한 역할을 하리라 추측된다.다.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1724-1730
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• 2012

An ${\alpha}$-L-arabinofuranosidase (TmAFase) from Thermotoga maritima MSB8 is a highly thermostable exo-acting hemicellulase that exhibits a relatively higher activity towards arabinan and arabinoxylan, compared with other glycoside hydrolase 51 family enzymes. In the present study, we carried out the enzymatic characterization and structural analysis of TmAFase. Tight domain associations found in TmAFase, such as an inter-domain disulfide bond (Cys306 and Cys476) in each monomer, a novel extended arm (amino acids 374-385) at the dimer interface, and total 12 salt bridges in the hexamer, may account for the thermostability of the enzyme. One of the xylan binding determinants (Trp96) was identified in the active site, and a region of amino acids (374-385) protrudes out forming an obvious wall at the substrate-binding groove to generate a cavity. The altered cavity shape with a strong negative electrostatic distribution is likely related to the unique substrate preference of TmAFase towards branched polymeric substrates.

• Molecules and Cells
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• 제46권12호
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• pp.764-777
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• 2023

Recombinant immunotoxins (RITs) are fusion proteins consisting of a targeting domain linked to a toxin, offering a highly specific therapeutic strategy for cancer treatment. In this study, we engineered and characterized RITs aimed at mesothelin, a cell surface glycoprotein overexpressed in various malignancies. Through an extensive screening of a large nanobody library, four mesothelin-specific nanobodies were selected and genetically fused to a truncated Pseudomonas exotoxin (PE24B). Various optimizations, including the incorporation of furin cleavage sites, maltose-binding protein tags, and tobacco etch virus protease cleavage sites, were implemented to improve protein expression, solubility, and purification. The RITs were successfully overexpressed in Escherichia coli, achieving high solubility and purity post-purification. In vitro cytotoxicity assays on gastric carcinoma cell lines NCI-N87 and AGS revealed that Meso(Nb2)-PE24B demonstrated the highest cytotoxic efficacy, warranting further characterization. This RIT also displayed selective binding to human and monkey mesothelins but not to mouse mesothelin. The competitive binding assays between different RIT constructs revealed significant alterations in IC₅₀ values, emphasizing the importance of nanobody specificity. Finally, a modification in the endoplasmic reticulum retention signal at the C-terminus further augmented its cytotoxic activity. Our findings offer valuable insights into the design and optimization of RITs, showcasing the potential of Meso(Nb2)-PE24B as a promising therapeutic candidate for targeted cancer treatment.

• Journal of Plant Biotechnology
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• 제42권4호
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• pp.277-283
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• 2015

Plant genome analyses, including Arabidopsis thaliana showed a large gene family of plant receptor kinases with various extracellular ligand-binding domain. Now intensively studies to understand physiological and cellular functions for higher plant receptor kinases in diverse and complex biological processes including plant growth, development, ligands perception including steroid hormone and plant-microbe interactions. Brassinosteroids (BRs) as a one of well know steroid hormone are plant growth hormones that control biomass accumulation and also tolerance to many biotic and abiotic stress conditions and hence are of relevance to agriculture. BRI1 receptor kinase, which is localized in plasma membrane in the cell sense BRs and it bind to a receptor protein known as BRASSINOSTEROID INSENSITIVE 1 (BRI1). Recently, we reported that BRI1 and its co-receptor, BRI1-ASSOCIATED KINASE (BAK1) autophosphorylated on tyrosine residue (s) in vitro and in vivo and thus are dual-specificity kinases. Other plant receptor kinases are also phosphorylated on tyrosine residue (s). Post-translational modifications (PTMs) can be studied by altering the residue modified by directed mutagenesis to mimic the modified state or to prevent the modification. These approaches are useful to not only characterize the regulatory role of a given modification, but may also provide opportunities for plant improvement.

• 한국자기공명학회논문지
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• 제15권1호
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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.

• 농촌지도와개발
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• 제22권3호
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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.