• 제목/요약/키워드: active site

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Bacillus Pumilus TX703 유래 Xylanase의 활성에 관여하는 아미노산 잔기의 확인 (Identification of Amino Acid Residues Involved in Xylanase Activity from Bacillus Pumilus TX703)

  • 박영서
    • 생명과학회지
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    • 제15권4호
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    • pp.633-640
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    • 2005
  • Bacillus pumilus TX703으로부터 xylanase를 정제한 후 효소의 활성부위를 조사하기 위하여 여러 가지 화학수식제를 사용하여 효소활성의 저해도를 측정하였다. 여러 가지 화학수식제 중에서 carbodiimide와 N-bromosuccinimide가 효소활성을 완전히 저 해시 켜 glutamic acid 또는 aspartic acid 잔기와 tryptophan 잔기가 효소의 활성부위에 관여하리라 추측되었다. 각각의 경우에 효소 실활은 수식제의 첨가농도에 따라 pseudo first-order kinetics 양식을 보여주었으며, car-bodiimide와 N-bromosuccinimide는 각각 비경쟁적 저해와 경쟁적 저해방식을 나타내었다. 기질첨가에 의한 효소활성 보호실험을 통하여 tryptophan 잔기가 기질결합부위라 판단되었다. 효소실활속도의 분석에 의해 효소활성에는 2개의 glutamic acid 또는 aspartic acid 잔기와 1개의 tryptophan 잔기가 관여하는 것으로 나타났다.

Electrochemical Studies of Immobilized Laccases on the Modified-Gold Electrodes

  • Yoon Chang-Jung;Kim Hyug-Han
    • 전기화학회지
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    • 제7권1호
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    • pp.26-31
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    • 2004
  • The direct electrochemical studies of four laccases (plant and fungal laccases) have been investigated on a gold electrode functionalized with a new tether of 2.2'-dithiosalicylic aldehyde. Results from these studies indicate that the redox potential of the active site of plant laccase from Rhus vernificera is shifted to a more negative value(255 mV versus SCE) than that of fungal laccase from Pyricularia oryzae (480 mV versus SCE). Mechanistic studies indicate that the reduction of type-1 Cu precedes the reduction of type-2 and type-3 Cu ions when the electrode is poised initially at different potentials. Also a new tether, 2.2'-dithiosalicylic aldehyde, has been used to study the redox properties of two laccases (LCCI and Lccla) covalently attached to a gold electrode. An irreversible peak at 0.47V vs. SCE is observed in the cyclic voltammorams of LCCI. In contrast, the cyclic voltammograms of LCCIa contain a quasi-reversible peak at 0.18V vs. SCE and an irreversible peak at 0.50V vs. SCE. We find that the replacement of the eleven amino acids a the C-terminus with a single cysteine residue $(i.e., \;LCCI{\rightarrow}LCCIa)$ influences the rate of heterogeneous electron transfer between an electrode and the copper containing active sites $(K_{het}\;for\;LCCI=1.0\times10^{-2}\;s^{-1}\;and\;K_{het}\;for\;LCCI_a= 1.0\;times10^{-1}\;s^{-1}\'at\;0.18V\;versus\;SCE\;and\;4.0\times10^{-2}\;s^{-1}\;at\;0.50V\; versus\;SCE)$. These results show for the first time that the change of the primary structure of a protein via site-directed mutagenesis influences both the redox potentials of the copper ions in the active site and the rate of heterogeneous electron transfer.

EMPLUX Collector에 의한 토양 오염 가스 조사 (Soil-Vapor Survey on Soil-Remediation by EMPLEX Collector)

  • 김정성
    • 한국환경과학회지
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    • 제17권1호
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    • pp.119-128
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    • 2008
  • Laboratory analytical results of 22 sets of hydrophobic adsorbent coils containing surface soil-vapor and two soil samples collected by conventional intrusive method from each boring location at two active dry cleaning facilities in the State of Illinois, U.S.A, were presented to evaluate the performance of soil-vapor survey. The most critical factor to determine the effectiveness of soil-vapor survey is the distance from the soil-vapor sampling device to the actual contamination, which is a function of soil porosity, permeability, primary lithology, and other geological and hydrogeological site-specific parameters. Also this factor can be affected by the history of contaminant-generating operations. The laboratory analytical results in this study showed longer dry cleaning operation history (i.e., 50 years) and presence of fine sand at the beneath Site B allow the contaminants to migrate farther and deeper over a fixed time compared to Site A(i.e., 35 years and silty clay) so that the soil-vapor survey is not likely the most effective environmental site investigation method alone for Site B. However, for Site A, the soil-vapor survey successfully screened the site to identify the location reporting the highest soil concentration of chlorinated solvents.

Biochemical Properties of Second Site Mutation of Human Immunodeficiency Virus Integrase

  • Kim, Do-Jin;Oh, You-Take;Lee, Sang-Kwang;Shin, Cha-Gyun
    • BMB Reports
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    • 제32권6호
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    • pp.599-604
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    • 1999
  • A highly conserved amino acid, glutamic acid (Glu), present at position 152 in the catalytic domain of the human immunodeficiency virus type 1 (HIV-1) integrase (IN) protein has been known to be critical for enzymatic function since substitution of Glu 152 with other residues results in a complete loss of enzymatic activities. In order to better understand the role of Glu 152 as a conserved residue in enzymatic action, intragenic second site mutations have been introduced around residue 152 of a mutant IN (E152A), and their biochemical properties were analyzed in terms of enzymatic activities. Disintegration activities were found to be significantly restored in several second site mutant INs, while integration activities were only recovered weakly. However, endonucleolytic activities were not discovered in all the mutant INs. These findings indicate that the second site mutations can partially restore that catalytic structure of the active site disturbed by the E152A mutation and lead to the regaining of integration and disintegration activities. In addition, it is also suggested that endonucleolytic activity requires a more accurate structure of the catalytic site than that for the integration and disintegration activities.

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Determination of Translocation and Deacylation Rate Constants for Complex Formation between Serpin and Protease

  • Shin, Jong-Shik;Yu, Myeong-Hee
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2001년도 학술 발표회 진행표 및 논문초록
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    • pp.62-62
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    • 2001
  • Serpins inhibit target proteases by forming tight acyl complex Via incorporation of the reactive center loop into ${\beta}$-sheet A. Metastability of the serpins control the translocation of the protease from the initial binding site to the opposite pole of the serpin. Recently the crystal structure of a serpin-protease complex revealed that the active site of the protease is distorted.(omitted)

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도시안전맵 웹사이트 구축을 위한 기존 웹사이트 분석연구 (Research for Building an Existing Web Site to Web Site Analysis of Urban Safety Map)

  • 임동현;김현수;김진욱;강부성
    • 한국주거학회:학술대회논문집
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    • 한국주거학회 2008년 추계학술발표대회 논문집
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    • pp.222-225
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    • 2008
  • In urban life, an interest in safety and demands for safety information about crimes and traffic are increasing. As well, communication on the internet is so active. In this study, we analyze web sites in US, UK, and JP, which have started safety map services, and then we provide basic material for the introduction of safety map into Korea. According to the analysis result, in US and UK, the safety map web sites are administered by the government and provided to the general public. The geographical area is provided around administrative districts and statistical data. In JP, The safety map web site provide not a one sided service but the service that user can directly take a part in, and also there is an web site for crimes against vulnerable people.

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중대재해처벌법 핵심 안전조항에 대한 건설업 종사자의 인식 고찰 (A Study on the Perception of Construction Workers on the Key Safety Provisions of the Serious Disaster Punishment Act)

  • 김재민
    • 대한안전경영과학회지
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    • 제24권4호
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    • pp.71-83
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    • 2022
  • This study is to investigate the degree of field awareness of how each provision of the Serious Disaster Punishment Act will affect accident prevention. As a result of conducting a survey of construction site workers, it was analyzed that construction site workers had a low sense of safety and did not voluntarily engage in safety activities. And it can be seen that they are taking a very passive position that it is efficient to implement safety under the supervision of the supervisor. Workers who need voluntary construction site safety activities are most aware of the situation at the site, but the problem was pointed out as "lack of procedures for listening to opinions and preparing improvement measures" for these problems. Future research is needed to see if this is a problem for individual workers or for those in charge of safety and health who do not correct it even though active opinions have been submitted.

Statistical Model to Describe Boiling Phenomena for High Heat Flux Nucleate Boiling and Critical Heat Flux

  • Ha, Sang-Jun;No, Hee-Cheon
    • 한국원자력학회:학술대회논문집
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    • 한국원자력학회 1996년도 추계학술발표회논문집(1)
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    • pp.230-235
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    • 1996
  • The new concept of dry area formation based on Poisson distribution of active nucleation sites and the concept of the critical active site density is presented. A simple statistical model is developed to predict the change of slope of the boiling curve up to critical heat flux (CHF) quantitatively. The predictions by the present model are in good agreement with the experimental data. Also it turns out that the present model well explains the mechanism on how the surface wettability influences CHF.

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Thioredoxin System and Redox Signaling; Defence against Stress and Toxicity

  • Yodoi, Junji;Masutani, Hiroshi;Nakamura, Hajime
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2001년도 International Symposium on Signal transduction in Toxicology
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    • pp.84-88
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    • 2001
  • Human Thioredoxin (TRX) with with redox-active dithiol (C-C-Y-C-) in the active site has been cloned as adult T cell leukemia derived factor produced by HTLV-I transformed cells. Thioredoxin (TRX) is one of the major components of the thiol-reducing system and plays multiple roles in cellular processes such as proliferation, apoptosis and gene expression.(omitted)

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Cytochrome c Peroxidase: A Model Heme Protein

  • Erman, James E.;Vitello, Lidia B.
    • BMB Reports
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    • 제31권4호
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    • pp.307-327
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    • 1998
  • Cytochrome c peroxidase (CcP) is a yeast mitochondrial enzyme which catalyzes the reduction of hydrogen peroxide to water using two equivalents of ferrocytochrome c. The CcP/cytochrome c system has many features which make it a very useful model for detailed investigation of heme protein structure/function relationships including activation of hydrogen peroxide, protein-protein interactions, and long-range electron transfer. Both CcP and cytochrome c are single heme, single subunit proteins of modest size. High-resolution crystallographic structures of both proteins, of one-to-one complexes of the two proteins, and a number of active-site mutants are available. Site-directed mutagenesis studies indicate that the distal histidine in CcP is primarily responsible for rapid utilization of hydrogen peroxide implying significantly different properties of the distal histidine in the peroxidases compared to the globins. CcP and cytochrome c bind to form a dynamic one-to-one complex. The binding is largely electrostatic in nature with a small, unfavorable enthalpy of binding and a large positive entropy change upon complex formation. The cytochrome c-binding site on CcP has been mapped in solution by measuring the binding affinities between cytochrome c and a number of CcP surface mutations. The binding site for cytochrome c in solution is consistent with the crystallographic structure of the one-to-one complex. Evidence for the involvement of a second, low-affinity cytochrome c-binding site on CcP in long-range electron transfer between the two proteins is reviewed.

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