• Title/Summary/Keyword: 기질상호작용

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Biochemical Characterization of Heterologously Expressed Chitinase 1 (Chi1) from an Inky Cap, Coprinellus congregatus (이형 재조합한 먹물버섯 Coprinellus congregatus Chitinase 1 (Chi1)의 발현과 생화학적 특성 분석)

  • Yoo, Yeeun;Choi, Hyoung T.
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.309-312
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    • 2013
  • Fungal cell walls consist of various glucans and chitin. Fungi produce chitinases for their growth and development. The inky cap, Coprinellus congregatus, produces at least two different chitinases during its life cycle. Chitinase 1 (chi1) is expresses throughout its life cycle while chitinase 2 (chi2) is expressed at the mushroom autolysing phase. The cloned cDNA of chi1 is successfully expressed as a fusion protein with c-myc in Pichia pastoris, and purified by the affinity chromatography. The optimum pH and temperature of Chi1 was pH 8.0 and $35^{\circ}C$, respectively when 4-nitrophenyl N,N',N"-triacetyl-${\beta}$-D-chitotrioside was used as the substrate. The $K_m$ value and $V_{max}$ for the substrate was 0.780 mM and 0.10 OD $min^{-1}unit^{-1}$, respectively. The addition of purified Chi1 resulted in total growth inhibition against several plant pathogenic fungi such as Alternaria alternata, Fusarium graminearum and Trichoderma harzianum at the concentration of 60 ${\mu}g/ml$.

3D-QSARs analyses for Tyrosinase Inhibitory Activity of 2-Phenyl-1,4-benzopyrone (Flavones) Analogues and Molecular Docking (2-Phenyl-1,4-benzopyrone 유도체 (Flavones)의 Tyrosinase 저해활성에 관한 3D-QSARs 분석과 분자도킹)

  • Park, Joon-Ho;Sung, Nack-Do
    • Journal of Applied Biological Chemistry
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    • v.53 no.4
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    • pp.225-231
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    • 2010
  • To understand the inhibitory activity with changing hydroxyl substituents ($R_l-R_9$) of polyhydroxy substituted 2-phenyl-l,4-benzopyrone analogues (1-25) against tyrosinase (PDB ID: oxy-form; 1WX2), molecular docking and the three dimensional quantitative structure-activity relationships (3D-QSARs: Comparative molecular field analysis (CoMFA) & Comparative molecular similarity indices analysis (CoMSIA)) were studied quantitatively. The statistically best models were CoMFA 1 and CoMSIA 1 model from the results. The optimized CoMSIA 1 model with the sensitivity of the perturbation and the prediction produced ($dq^2'/dr_{yy'}^2$=1.009 & $q^2$=0.51l) by a progressive scrambling analysis were not dependent on chance correlation. The inhibitory activities with optimized CoMSIA 1 model were dependent upon electrostatic factor (51.4%) of substrate molecules. Contour mapping the 3D-QSAR models to the active site of tyrosinase provides new insight into the interaction between tyrosinase as receptor and 2-phenyl-l,4-benzopyrone analogues as inhibitor. Therefore, the results will he able to apply to the optimization of a new potent tyrosinase inhibitors.

2D-QSAR and HQSAR Analysis on the Herbicidal Activity and Reactivity of New O,O-dialkyl-1-phenoxy-acetoxy-1-methylphosphonate Analogues (새로운 O,O-dialkyl-1-phenoxyacetoxy-1-methylphosphonate 유도체들의 반응성과 제초활성에 관한 2D-QSAR 및 HQSAR 분석)

  • Sung, Nack-Do;Jang, Seok-Chan;Hwang, Tae-Yeon
    • The Korean Journal of Pesticide Science
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    • v.11 no.2
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    • pp.72-81
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    • 2007
  • Quantitative structure-activity relationships (QSARs) on the pre-emergency herbicidal activity and reactivity of a series of new O,O-dialkyl-1-phenoxyacetoxy-1-methylphosphonates (S) analogues against seed of cucumber (Cucumus Sativa) were discussed quantitatively using 2D-QSAR and HQSAR methods. The statistical values of HQSAR model were better than that of 2D-QSAR model. From the frontier molecular orbital (FMO) interaction between substrate molecule (S) and $BH^+$ ion (I) in PDH enzyme, the electrophilic reaction was superior in reactivity. From the effect of substituents, $R_2$-groups in substrate molecule (S) contributed to electrophilic reaction with carbonyl oxygen atom while X, Y-groups contributed to nucleophilic reaction with carbonyl carbon atom. And the influence of X,Y-groups was more effective than that of $R_2$-groups. As a results of 2D-QSAR model (I & II) and atomic contribution maps with HQSAR model, the more length of X, Y-groups is longer, the more herbicidal activity tends to increased. And also, the optimal ${\epsilon}LUMO$ energy, $({\epsilon}LUMO)_{opt.}$=-0.479 (e.v.) of substrate molecule is important factor in determining the herbicidal activity. It is predicted that the herbicidal activity proceeds through a nucleophilic reaction. From the analytical results of 2D-QSAR and HQSAR model, it is suggested that the structural distinctions and descriptors that contribute to herbicidal activities will be able to applied new herbicide design.

CHILDHOOD TRAUMA:RESILIENCE AND RISK FACTORS ON DEVELOPMENTAL TRAJECTORY (소아기 외상 : 발달경로에 따른 보호 및 위험인자)

  • Kim, Young-Shin
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.13 no.1
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    • pp.15-23
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    • 2002
  • Knowledge regarding the resilience factors and risk factors of the childhood trauma on the developental trajectory is in its infancy due to the lack of prospective follow-up studies in the childhood trauma and limited understanding of the complex reciprocal interactions between childhood trauma, develop-ent and various aspects of children's environment. These difficulties in the conceptual framework and research methods in the childhood trauma are partly reflected in the inconsistencies, even controversies, of the results in the childhood trauma researches. Despite these difficulties, common aspects of the risk factors and resilience of the childhood trauma on the development can be identified from the previous studies. The resilience to the negative outcome on the development by childhood trauma includes:sex female before puberty, male after puberty or infancy), high socioeconomic status, no organic problem, easy temperament, no previous experience with early loss or separation, younger age at the trauma, better problem solving capacity, high self-esteem, internal locus of control, high coping skills, ability to identify interpersonal relationships, ability to play, sense of humor, having capable parents, having a warm relaionship with at least one of the parents, high education and participating in the organized religious activities. These commonalities of the results suggest that risk and resilient factors of the childhood trauma are interdependent, each factor has multiplicity in the impacts on the children's development according to the developmental stage of the child, family and children's other environment, trauma and stressor have diverse effects according to their intensity and risk and resilience factors could have synergistic or antagonistic effects to each other. To develop comprehensive understanding on the relationship between childhood trauma and developmental psychopathology, risk and resilience factors and to develop effective and efficient prevention and intervention, research on the effect of the stress on the neurodevelopment, on the individual differences of the response to the trauma including genetic factors and constitution, and on the brain plasticity should be accompanied in the future.

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Electrostatic Immobilization of D-Xylose Isomerase to a Cation Exchanger for the Conversion of D-Xylose to D-Xylulose (D-xylose에서 D-xylulose로의 전환을 위한 D-xylose Isomerase의 정전기적 고정화)

  • Hang, Nguyen Thi;Kim, Sung-Gun;Kweon, Dae-Hyuk
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.163-167
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    • 2012
  • Since D-xylose is not fermentable in Saccharomyces cerevisiae, its conversion to D-xylulose is required for its application in biotechnological industries using S. cerevisiae. In order to convert D-xylose to D-xylulose by way of an enzyme immobilized system, D-xylose isomerase (XI) of Escherichia coli was fused with 10-arginine tag (R10) at its C-terminus for the simple purification and immobilization process using a cation exchanger. The fusion protein XIR10 was overexpressed in recombinant E. coli and purified to a high purity by a single step of cation exchange chromatography. The purified XIR10 was immobilized to a cation exchanger via the electrostatic interaction with the C-terminal 10-arginine tag. Both the free and immobilized XIR10 exhibited similar XI activities at various pH values and temperatures, indicating that the immobilization to the cation exchanger has a small effect on the enzymatic function of XIR10. Under optimized conditions for the immobilized XIR10, D-xylose was isomerized to D-xylulose with a conversion yield of 25%. Therefore, the results of this study clearly demonstrate that the electrostatic immobilization of XIR10 via the interaction between the 10-arginine tag and a cation exchanger is an applicable form of the conversion of D-xylose to D-xylulose.

User-centric Context Model for Context-aware Application (맥락 인식 애플리케이션을 위한 사용자 중심의 맥락 모델)

  • Hong, Dong-Pyo;Woo, Woon-Tack
    • Proceedings of the Korea Information Processing Society Conference
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    • 2007.05a
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    • pp.810-813
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    • 2007
  • 본 논문에서는 맥락 인식 (context-aware) 애플리케이션 개발에 있어서 사용자의 맥락 정보를 보다 효과적으로 처리할 수 있는 사용자 중심의 맥락 모델을 제안한다. 유비쿼터스 컴퓨팅 개념의 확산과 함께 맥락 인식 애플리케이션들에 관한 많은 연구가 진행되고 있다. 하지만, 맥락에 대한 정의는 여전히 모호하며, 애플리케이션들 마다 서로 다른 형태의 맥락을 활용하고 있기 때문에, 맥락에 대한 보다 구체적인 정의와 다양한 애플리케이션에 활용 가능한 형태의 맥락 모델이 필요하다. 제안된 사용자 중심의 맥락 모델에서는 사용자가 애플리케이션과 상호작용할 때 사용자의 직접적인 명령을 제외한 사용자와 관련된 정보를 맥락으로 정의한다. 또한, 제안된 사용자 중심의 맥락은 5W1H 형태로 구조화한 맥락요소 (ContextElement), 맥락 요소들을 편리하게 처리할 수 있는 연산자들을 포함하는 맥락 (Context), 그리고 단편적인 맥락 정보뿐만 아니라 기존의 맥락 정보까지도 활용할 수 있는 맥락메모리 (ContextMemory)로 구성된다. 특히, 다양한 센서들로부터 획득된 정보를 맥락 모델의 인터페이스를 통해서 맥락 인식 애플리케이션에서 활용할 수 있기 때문에, 서로 다른 맥락 인식 애플리케이션들을 개발함에 있어서도 동일한 맥락 모델을 사용할 수 있는 장점이 있다. 제안된 맥락 모델의 유용함을 보이기 위해서, 센서로부터 획득된 맥락 정보를 처리하는데 소요되는 시간을 측정하는 실험을 하였다. 따라서 제안된 사용자 중심의 맥락 모델은 사용자와 맥락 인식 애플리케이션간 자연스러운 상호작용을 지원할 것으로 기대된다.) kcal/mol의 생성활성화 에너지 감을 나타내었고, TGA로부터의 분해활성화 에너지는 각각 31.94, 30.84, 24.16 kcal/mol의 값을 나타내었다.로 감소되었다(35.2% vs. 77.4%; p<0.01). 실험 2에서 다양한 정자 농도에 의한 정자 침투율과 정상 수정률을 바탕으로 판단했을 때 $4.6{\times}10^6/ml$의 정자 농도가 다른 정자 농도에 비해 난구 세포부착 난자의 체외 수정에 적합한 것으로 나타났다. 체외 수정과정에서 난구 세포 부착된 상태로 수정된 난자는 나화 난자에 비해 유의적으로(p<0.05) 높은 분할률(48.8% vs. 58.9%), 배반포 형성률(11.0% vs. 22.8%)과 배반포 세포수$(22{\pm}2\;vs.\;29{\pm}2)$를 나타내었다. 본 연구의 결과로부터 돼지의 체외 수정과정에서 난구 세포의 존재는 정자 침투를 저해하지만 분할률, 배반포 형성률 및 배반포의 세포수를 증가시키는 것으로 사료된다.수의 유출입 지점에 온도센서를 부착하여 냉각수의 온도를 측정하고 냉각수의 공급량과 대기의 온도 등을 측정하여 대사열의 발생을 추정할 수 있었다. 동시에 이를 이용하여 유가배양시 기질을 공급하는 공정변수로 사용하였다 [8]. 생물학적인 폐수처리장치인 활성 슬러지법에서 미생물의 활성을 측정하는 방법은 아직 그다지 개발되어있지 않다. 본 연구에서는 슬러지의 주 구성원이 미생물인 점에 착안하여 침전시 슬러지층과 상등액의 온도차를 측정하여 대사열량의 발생량을 측정하고 슬러지의 활성을 측정할 수 있는 방법을 개발하였다.enin과

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Site-directed Mutagenesis Analysis Elucidates the Role of 223/227 Arginine in 23S rRNA Methylation, Which Is in 'Target Adenine Binding Loop' Region of ErmSF (위치 지정 치환 변이를 이용한 ErmSF의 '타깃 Adenine Binding Loop'을 형성하는 부위에 존재하는 223/227 Arginine 잔기의 23S rRNA Methylation 활성에서의 역할 규명)

  • Jin, Hyung-Jong
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.79-86
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    • 2012
  • ErmSF is one of the Erm family proteins which catalyze S-adenosyl-$_L$-methionine dependent modification of a specific adenine residue (A2058, E. coli numbering) in bacterial 23S rRNA, thereby conferring resistance to clinically important macrolide, lincosamide and streptogramin B ($MLS_B$) antibiotics. $^{222}FXPXPXVXS^{230}$ (ErmSF numbering) sequence appears to be a consensus sequence among the Erm family. This sequence was supposed to be involved in direct interaction with the target adenine from the structural studies of Erm protein ErmC'. But in DNA methyltarnsferase M. Taq I, this interaction have been identified biochemically and from the complex structure with substrate. Arginine 223 and 227 in this sequence are not conserved among Erm proteins, but because of the basic nature of residues, it was expected to interact with RNA substrates. Two amino acid residues were replaced with Ala by site-directed mutagenesis. Two mutant proteins still maintained its activity in vivo and resistant to the antibiotic erythromycin. Compared to the wild-type ErmSF, R223A and R227A proteins retained about 50% and 88% of activity in vitro, respectively. Even though those arginine residues are not essential in the catalytic step, with their positive charge they may play an important role for RNA binding.

A Study on the Career Decision-making Process of Female Undergraduates who Want the Male-dominated Job: A Grounded Theory Based Approach (남성중심적 직업을 희망하는 여자대학생들의 직업결정과정: 근거이론 접근)

  • Kim, Young Shil;Lim, Sung Moon
    • The Korean Journal of Woman Psychology
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    • v.16 no.3
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    • pp.303-329
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    • 2011
  • In Korea, recent women's educational standard is more higher than any other period but rate of using human resources is the lowest level among the OECD nations. As the reason of this phenomenon, the career choice attract attention. Traditionally, job is classified female-dominated job from male-dominated job, and most women still avoid male-dominated job. If this unequal concentrated phenomenon is relieved, diversity using of human resource will be available. For the purpose, this study try to determine the experience structure and the career decision-making process of the female undergraduate who want the male-dominated job through the qualitative study method, and to determine factors that work negative or positive on the process to use for the career consult. Grounded theory based data analyses resulted in 61 concepts, 27 sub-categories, and 12 categories from the recorded data of 8 female undergraduates who want the male-dominated job. The core category was "For the special life, keeping challenging with the will". A paradigm model consist of the casual condition, central phenomenon, contextual condition, intervening condition, actions/reactions and the result was proposed. And the participants went through five stages with time in their career decision-making process: the stage of awareness about the trait and aspiration, the stage of internal determination, the stage of support and accept, the stage of challenge and overcome, and the stage of affirmation and confidence. Implications of the findings and suggestions for the future research are discussed.

Lipoxygenase Activity of Milled Fractions from Brown Rice (현미 도정획분의 Lipoxygenase 활성)

  • Kim, Ki-Joong;Rhee, Chong-Ouk
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.145-149
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    • 1997
  • Lipoxygenase activity from brown rice varieties (Tongjinbyeo, Kumohbyeo and Kanchukbyeo) was investigated using spectrophotometric method. In all three varieties, there was an increase in the enzyme activity with the reaction time. Enzyme activity was tested at different concentration of the substrate. The $V_{max}\;and\;K_m$ values of Tongjin, Kumoh and Kanchukbyeo were 57.89, 19.85 and 31.38 units/mg protein and 0.054, 0.045 and 0.035 mM. The study of lipoxygenase activity at different pH levels showed that all the varieties had maximal activities around $pH\;7.0{\sim}7.6$. The enzyme activity and specific activity on milled fractions of different brown rice varieties, fraction II was superior to the other fractions and fraction IV was inferior to the other fractions. As the result of microwave heating for 0, 30, 60 and 90 sec, the enzyme activity and specific activity of all the varieties were decreased by the elapse of heating time.

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THE ADHESION OF ODONTOBLAST TO TYPE I COLLAGEN (상아모세포의 I 형 아교질에 대한 부착)

  • Ahn, Myung-Ki;Jeong, Tae-Sung;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.3
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    • pp.308-316
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    • 2010
  • Odontoblasts are anchorage dependent cells adhering to a substrate via cell adhesive molecules. Receptor ligands such as integrins bind to these proteins and are known to function as signal transduction molecules in a series of critical recognition events of cell-substratum. The aim of this study is to examine the interaction of odontoblast (MDPC-23 cell) with type I Col and the effect of TGF-${\beta}1$ and TNF-$\alpha$ on the expression of cell adhesion molecules. In this study, MDPC-23 cells adhered to type I Col dose-dependently. Immunofluorescence data demonstrated that integrin ${\alpha}1$, ${\alpha}2$ and CD44 were expressed on cell surface, and FAK and paxillin were localized in focal adhesion plaques in MDPC-23 cells adhesion to Col. Cytokine TGF-${\beta}1$ increased the adhesion of MDPC-23 cells to Col and the expression level of integrin ${\alpha}1$, 4{\alpha}2$ and chondroitin sulfate on MDPC-23 cells. RT-PCR data demonstrated that cytokine TGF-${\beta}1$ increased the amount of integrin ${\alpha}1$ mRNA in MDPC-23 cells. Therefore, MDPC-23 cells adhere to collagen type I Col and expressed a complex pattern of integrins and proteoglycans, including ${\alpha}1$, ${\alpha}2$, chondroitin sulfate and CD44 detected by immunoblotting and immunofluorescence assay. TGF-${\beta}1$ treatment enhanced the expression of adhesion molecules such as integrin ${\alpha}1$, ${\alpha}2$ and chondroitin sulfate.