• BMB Reports
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• 제41권5호
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• pp.353-357
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• 2008

LRR family proteins play important roles in a variety of physiological processes. To facilitate their production and crystallization, we have invented a novel method termed "Hybrid LRR Technique". Using this technique, the first crystal structures of three TLR family proteins could be determined. In this review, design principles and application of the technique to protein crystallization will be summarized. For crystallization of TLRs, hagfish VLR receptors were chosen as the fusion partners and the TLR and the VLR fragments were fused at the conserved LxxLxLxxN motif to minimize local structural incompatibility. TLR-VLR hybridization did not disturb structures and functions of the target TLR proteins. The Hybrid LRR Technique is a general technique that can be applied to structural studies of other LRR proteins. It may also have broader application in biochemical and medical application of LRR proteins by modifying them without compromising their structural integrity.

• Journal of Microbiology and Biotechnology
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• 제9권4호
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• pp.381-385
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• 1999

Taq DNA polymerase from Thermus aquaticus has been shown to be very useful in a polymerase chain reaction. Taq DNA polymerase has a domain at the amino terminus (residues 1 to 290) that has 5'-3' exonuclease activity and a domain at the C-terminus that catalyzes the polymerase reaction. Taq DNA polymerase is classified into the Pol I family, which is represented by E. coli DNA polymerase I. The alignment of amino acid sequences for the 5'-3' exonuclease domains of the Pol I family DNA polymerases shows ten highly conserved carboxylic amino acids. Crystallographic studies suggested that six of the carboxylic amino acids are clustered within a 7 $\AA$ radius by chelating three metal ions in the active site. Those six carboxylic residues are mutagenized to alanines in order to better understand their function. All six carboxylic residues, Asp l8, Glu1l7, Asp1l9, Asp120, Asp142, and Aspl44, are crucial for catalysis of 5'-3' exonuclease.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.33-33
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• 2003

Human DJ-1 and Escherichia coli Hsp31 belong to ThiJ/PfpI family whose members contain a conserved domain. DJ-1 is associated with autosomal recessive early-onset parkinsonism and Hsp31 is a molecular chaperone. Structural comparisons between DJ-1, Hsp31, and an archeal protease, a member of ThiJ/PfpI family, lead to the identification of the chaperons activity of DJ-1 and the proteolytic activity of Hsp31. Moreover, the comparisons provide insights into how the functional diversity is realized in proteins that share an evolutionarily conserved domain. On the basis of the chaperons activity, the possible role of DJ-1 in the pathogenesis of Parkinson's disease is discussed.

• Journal of Industrial and Engineering Chemistry
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• 제67권
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• pp.293-300
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• 2018

Lipocalins are diverse group of small extracellular proteins found in various organisms. In this study, members of 10 non-homologous lipocalin-ligand crystal complex structures were remodeled using rigid and flexible ligand modes to validate the prediction efficiency of molecular docking simulation. The modeled ligand conformations indicated a high prediction accuracy in rigid ligand mode using cluster based analysis for most cases whereas the flexible ligand mode required further considerations such as ligand binding energy and RMSD for some cases. This in silico study is expected to serve as a platform in the screening of novel ligands against lipocalin family of proteins.

• BMB Reports
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• 제33권1호
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• pp.89-91
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• 2000

In this investigation we report our search for the presence of Leucine Rich Repeats (LRRs) in various Bacillus thuringiensis (Bt) sub species. Leucine rich repeats are short sequence motifs present in some proteins. The consensus sequence corresponding to the LRR was present in Crystal proteins of Bacillus thuringiensis sub species. This LRR sequence has been predicted to be involved in proteinprotein interactions or receptor binding functions, hence the importance of this study.

• Toxicological Research
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• 제18권2호
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• pp.183-190
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• 2002

The mechanism by which catechin-mediated cytotoxicity against tumor cells remains to be elusive. To elucidate the mechanical mights of anti-tumor effects, (-)epigallocatechin-gallate (EGCG) of catechin was applied to human prostate cancer DU 145 cells. Cell viability was measured by crystal violet staining. Cell lysates were wed to measure the catalytic activity of caspases by using fluorogenic peptide: Ac-DEVD-AMC for caspase-3 protease, Z-IETD-AFC for caspase-8 protease, Ac-LEHD-AFC for caspase-9 protease as substrates. The equal amounts of protein from cell lysate was separated on SDS-PAGE and analyzed by western blotting with anti-Fas antibody, anti-FasL antibody, anti-BCL2 antibody and anti-Bax antibody. (-)EGCG induced the death of DUl45 cells, which was revealed as apoptosis shown by DNA fragmentation. (-)EGCG induced the activation of caspase family cysteine proteases including caspase-3, -8 and -9 proteases in DU145 cells. Also, (-)EGCG increased the expression of Fas and Fas ligand (FasL) protein in DU145 colls. The expression level of BCL2 was decreased in (-)EGCG treated DU145 cells, whereas Bax protein was increased in a time-dependent manner. We suggest that (-)EGCG-induced apoptosis of DU145 cells is mediated by signaling pathway involving caspase family cysteine protease, mitochondrial BCL2-family protein and Fas/FasL.

• 한국방사선학회논문지
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• 제5권6호
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• pp.357-361
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• 2011

본 연구에서는 II-IV 족 화합물 반도체인 CdS를 이용한 액정광변조 방식의 X-ray 검출 시스템을 제안하였다. 제안된 시스템은 검출부, 신호처리부, 액정 구동 및 투과량 측정부, 마이크로컨트롤러부, 입출력부로 구성되었으며, 소형화 및 휴대형에 적합하게 제작되었다. 또한, 검출 범위 선택을 통하여 광범위한 조건에서 측정이 가능하도록 구성하였다. 제안된 시스템의 성능을 평가하기 위하여 조사선량 변화에 따른 CdS 센서의 출력 특성을 확인하였으며, 우수한 상관관계를 확인할 수 있었다. 또한, 인가전압에 따른 액정의 변화를 관찰하여 인가 전압에 따른 광투과율을 측정하였으며, 높은 상관관계와 우수한 재현성을 확인할 수 있었다. 이러한 결과를 통하여 본 연구에서 제안된 액정 광변조 방식의 특징인 우수한 재현성과 노이즈 내성 특성을 확인할 수 있었으며, 본 연구를 통하여 제안된 CdS 셀 기반 광변조 방식의 휴대형 X선 검출 시스템이 소형, 저가형, 휴대형 시스템으로 적용이 가능할 것으로 판단되었다.

• Molecules and Cells
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• 제38권2호
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• pp.105-111
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• 2015

The beta2-adrenergic receptor (${\beta}2AR$) belongs to the G protein coupled receptor (GPCR) family, which is the largest family of cell surface receptors in humans. Extra attention has been focused on the human GPCRs because they have been studied as important protein targets for pharmaceutical drug development. In fact, approximately 40% of marketed drugs directly work on GPCRs. GPCRs respond to various extracellular stimuli, such as sensory signals, neurotransmitters, chemokines, and hormones, to induce structural changes at the cytoplasmic surface, activating downstream signaling pathways, primarily through interactions with heterotrimeric G proteins or through G-protein independent pathways, such as arrestin. Most GPCRs, except for rhodhopsin, which contains covalently linked 11 cis-retinal, bind to diffusible ligands, having various conformational states between inactive and active structures. The first human GPCR structure was determined using an inverse agonist bound ${\beta}2AR$ in 2007 and since then, more than 20 distinct GPCR structures have been solved. However, most GPCR structures were solved as inactive forms, and an agonist bound fully active structure is still hard to obtain. In a structural point of view, ${\beta}2AR$ is relatively well studied since its fully active structure as a complex with G protein as well as several inactive structures are available. The structural comparison of inactive and active states gives an important clue in understanding the activation mechanism of ${\beta}2AR$. In this review, structural features of inactive and active states of ${\beta}2AR$, the interaction of ${\beta}2AR$ with heterotrimeric G protein, and the comparison with ${\beta}1AR$ will be discussed.

• 통합자연과학논문집
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• 제3권2호
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• pp.72-77
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• 2010

Alginate lyases, also known as alginases or alginate depolymerases, catalyze the degradation of alginate by a ${\beta}$-elimination mechanism that has yet to be fully elucidated. Alginate is a copolymer of ${\alpha}$-L-guluronate (G) and its C5 epimer ${\beta}$-D-mannuronate (M), arranged as homopolymeric G blocks, M blocks, alternating GM or random heteropolymeric G/M stretches. Almost all alginate lyases depolymerize alginate in an endolytical fashion via a ${\beta}$-elimination reaction. The alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, consisting of 776 amino-acid residues, is a novel exotype alginate lyase classified into polysaccharide lyase family 15. Till now there is no crystal structure available for this class of proteins. Since there is no template with high sequence identity, three-dimensional coordinates for exotype alginate lyase (PL 15 family) were determined using modeling methods (Comparitive modeling and Fold recognition). The structures were modeled using the X-ray coordinates from Heparinase protein family (PDB code: 3E7J). This enzyme (Atu3025) displays enzymatic activity for both poly-M and poly-G alginate. Since poly-M is widespread; docking of a tri-mannuronate against the modeled structure was performed. We identified some of those residues which are crucial for lyase activity. The results from this study should guide future mutagenesis studies and also provides a starting point for further proceedings.

• 한국결정성장학회:학술대회논문집
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• 한국결정성장학회 1997년도 Proceedings of the 12th KACG Technical Meeting and the 4th Korea-Japan EMGS (Electronic Materials Growth Symposium)
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• pp.145-145
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• 1997

Recent progress of electric technology requires new piezoelectric crystals having superior properties such as zero temperature coefficients and large electromechanical coupling factors. We have developed a series of new leading chandidates, La$_3$Ga5SiO14(langasite, LGS), La3Nb0.5Ga5.5O14(LNG), La3Ta0.5O14(LTG), to satisfy those requirements. High quality LGS, LNG and LTG single crystals, with dimensions of 2 inches in diameter, were successfully grown by the Czochralski method at a pulling rate of 1mm/h. Since no variation of chemical composition was observed when whole melt in a crucible was crystallized, congruency of these compositions was confirmed. Physical constants such as elastic constants, dielectric constants and piezoelectric constants were measured. Filters and oscillators made of grown LGS, LNG and LTG single crystals showed superior properties such as three times wider passband than that of quartz, low insertion loss and easy processing, Langasite family crystals were shown to be superior materials to other known materials such as quartz, LiTaO$_3$, $\alpha$-AlPO$_4$ and Li$_2$B$_4$O7.