• 제목/요약/키워드: Activity Form

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Inactive but Dimeric Form of Lipoprotein Lipase in Human Plasma

  • Park, Byung-Hyun
    • BMB Reports
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    • 제34권4호
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    • pp.329-333
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    • 2001
  • Active lipoprotein lipase (LPL) is known as a noncovalent homodimer of identical subunits, and dissociation of the dimer to a monomeric form renders the lipase inactive. In this study, the oligomerization status of LPL in human and rat plasma was investigated. The LPL activity was barely detectable in the control rat and human plasma. After the injection of heparin, the total lipolytic activity of plasma was rapidly increased, and reached its maximum in 30 min. Changes of the LPL protein correlated well with those of lipolytic activity. The LPL protein that is released by heparin into both human and rat plasma was active and dimeric in the sucrose density gradient ultracentrifugation. In control rat plasma, LPL was inactive, and a great fraction was present as an aggregate. However, the inactive LPL protein in the control human plasma retained the dimeric state, indicating that dimerization can be an entity independent of the catalytic activity of LPL. The released LPL is transported as a complex with lipoproteins in plasma. Lipoprotein profiles, determined by NaBr ultracentrifugation, exhibited typical LDL- and HDL-mammal patterns in humans and rats, respectively, with a smaller amount of the LDL fraction observed in rats. The difference in the lipoprotein profiles might influence the fate of the released LPL in plasma.

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단백질의 구조 전환과 기능 조절 (Conformational Switch and Functional Regulation of Proteins)

  • 유명희
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2001년도 추계학술대회 논문집 Vol.14 No.1
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    • pp.3-6
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    • 2001
  • In common globular proteins, the native form is in its most stable state. However, the native form of inhibitory serpins (serine protease inhibitors) and some viral membrane fusion proteins is in a metastable state. Metastability in these proteins is critical to their biological functions. Our previous studies revealed that unusual interactions, such as side-chain overpacking, buried polar groups, surface hydrophobic pockets, and internal cavities are the structural basis of the native metastability. To understand the mechanism by which these structural defects regulate protein functions, cavity-filling mutations of a 1-antitrypsin, a prototype serpin, were characterized. Increasing conformational stability is correlated with decreasing inhibitory activity. Moreover, the activity loss appears to correlate with the decrease in the rate of the conformational switch during complex formation with a target protease. We also increased the stability of a 1-antitrypsin greatly via combining various stabilizing single amino acid substitutions that were distributed throughout the molecule. The results showed that a substantial increase of stability, over 13 kcal/mol, affected the inhibitory activity with a correlation of 11% activity loss per kcal/mol. The results strongly suggest that the native metastability of proteins is indeed a structural design that regulates protein functions and that the native strain of a 1-antitrypsin distributed throughout the molecule regulates the inhibitory function in a concerted manner.

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단백질의 구조 전환과 기능 조절 (Conformational Switch and Functional Regulation of Proteins)

  • 유명희
    • E2M - 전기 전자와 첨단 소재
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    • 제14권12호
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    • pp.3-6
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    • 2001
  • In common globular proteins, the native form is n its most stable state. However, the native form of inhibitory serpins (serine protease inhibitors) and some viral membrane fusion proteins is in a metastable state. Metastability in these proteins is critical to their biological functions. Our previous studies revealed that unusual interactions, such as side-chain overpacking, buried polar groups, surface hydrophobic pockets, ad internal cavities are the structural basis of the native metastability. To understand the mechanism by which these structural defects regulate protein functions, cavity-filling mutations of $\alpha$1-antitrypsin, a prototype serpin, were characterized. Increasing conformational stability is correlated with decreasing inhibitory activity. Moreover, the activity loss appears to correlate with the decrease in the rate of the conformational switch during complex formation with a target protease. We also increased the stability of $\alpha$1-antitrypsin greatly via combining various stabilizing single amino acid substitutions that were distributed throughout the molecule. The results showed that a substantial increase of stability, over 13 kcal/mol, affected the inhibitory activity with a correlation of 11% activity loss per kcal/mol. The results strongly suggest that the native metastability of proteins is indeed a structural design that regulates protein functions and that the native strain of $\alpha$1-antitrypsin distributed throughout the molecule regulates the inhibitory function in a concerted manner.

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단백질의 구조 전환과 기능 조절 (Conformational Switch and Functional Regulation of Proteins)

  • 유명희
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2001년도 추계학술대회 논문집
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    • pp.3-6
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    • 2001
  • In common globular proteins, the native form is in its most stable state. However, the native form of inhibitory serpins (serine protease inhibitors) and some viral membrane fusion proteins is in a metastable state. Metastability in these Proteins is critical to their biological functions. Our previous studies revealed that unusual interactions, such as side-chain overpacking, buried polar groups, surface hydrophobic pockets, and internal cavities are the structural basis of the native metastability. To understand the mechanism by which these structural defects regulate protein functions, cavity-filling mutations of ${\alpha}$1-antitrypsin, a prototype serpin, were characterized. Increasing conformational stability is correlated with decreasing inhibitory activity. Moreover, the activity loss appears to correlate with the decrease in the rate of the conformational switch during complex formation with a target protease. We also increased the stability of ${\alpha}$1-antitrypsin greatly via combining various stabilizing single amino acid substitutions that were distributed throughout the molecule. The results showed that a substantial increase of stability, over 13 kcal/mol, affected the inhibitory activity with a correlation of 11% activity loss per kcal/mol. The results strongly suggest that the native metastability of proteins is indeed a structural design that regulates protein functions and that the native strain of e 1-antitrypsin distributed throughout the molecule regulates the inhibitory function in a concerted manner.

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Processing of an Intracellular Immature Pullulanase to the Mature Form Involves Enzymatic Activation and Stabilization in Alkaliphilic Bacillus sp. S-1

  • Lee, Moon-Jo;Kang, Bong-Seok;Kim, Dong-Soo;Kim, Yong-Tae;Kim, Se-Kwon;Chung, Kang-Hyun;Kim, Jume-Ki;Nam, Kyung-Soo;Lee, Young-Choon;Kim, Cheorl-Ho
    • BMB Reports
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    • 제30권1호
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    • pp.46-54
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    • 1997
  • Alkaliphilic Bacillus sp. S-1 secretes a large amount (approximately 80% of total pullulanase activity) of an extracellular pullulanase (PUL-E). The pullulanase exists in two forms: a precursor form (PUL-I: $M_r$ 180,000), and a processed form (PUL-E: $M_r$ 140,000). Two forms were purified to homogeneity and their properties were compared. PUL-I was different in molecular weight, isoelectric point, $NH_2$-terminal amino acid sequence, and stabilities over pH and temperature ranges. The catalytic activities of PUL-I were also distinguishable in the $K_m$ and $V_{max}$ values for various substrates, and in the specific activity for pullulan hydrolysis. PUL-E showed 10-fold higher specific activities than PUL-I. However. PUL-I is immunologically identical to PUL-E, suggesting that PUL-I is initially synthesized and proteolytically processed to the mature form of PUL-E. Processing was inhibited by PMSF, but not by pepstatin, suggesting that some intracellular serine proteases could be responsible for processing of the PUL-I. PUL-I has a different conformational structure for antibody recognition from that of PUL-E. It is also postulated that the translocation of alkaline pullulanase(AP) in the bacterium possibly requires processing of the $NH_2$-terminal region of the AP protein. Processing of the precursor involves a conformational shift. resulting in a mature form. Therefore. precursor processing not only cleaves the signal peptide, but also induces conformational shift. allowing development of active form of the enzyme.

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목재변색균(木材變色菌) 및 표면오염균류(表面汚染菌類)에 대(對)한 Streptomyces rimosus의 항균대사(抗菌代謝) (I) -2차(次) 대사물질(代謝物質)의 항균효능(抗菌效能)- (Antifungal Metabolisms of Streptomyces rimosus against Sapstain and Mold Fungi(I) -Antifungal Efficacy of Secondary Metabolites-)

  • 강규영;이동흡;오정수
    • Journal of the Korean Wood Science and Technology
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    • 제23권1호
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    • pp.42-48
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    • 1995
  • The purpose of this study is to evaluate the efficacy of metabolites produced form Streptomyces rimosus in controlling the growth of sapwood - inhabiting fungi. In order to carry out this task, the following specific fungi were tested : sapstain fungi - Ceratocystis pilifera, Ceratocystis piceae, and Aureobasidium pullulans ; mold fungi - Trichoderma hazianum, Trichoderma viride, Penicillium cirtrinum, and Aspergillus niger. Based on the tests, the following observations can be drawn. 1. The conidial germination of sapstain and mold fungi was completely inhibited leaving a clear zone around the paper disc treated with metabolites. The best inhibition was observed in A. pullulans plate and the least in T. viride. 2. Concentration of SB medium for the production of metabolites from St, rimosus affected antifungal activity of metabolites against sapwood - inhabiting fungi. Metabolites prepared from 1/3${\times}$SB medium showed the best activity and the least activity was observed in metabolites form 1/4${\times}$medium. 3. in vivo and in vitro test using wood blocks, treatment of pine sapwood blocks with metabolites also inhibited conidial germination and thus prevented discoloration. 4. Treatment with metabolites did not change the macroscopic structure of wood and did not cause the discoloration of the surface of wood by pigments produced form St. rimosus. In conclusion the results of this study indicate that antifungal metabloites of St, rimosus could be used for the biological control of sapstain and mold fungi.

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전신형태 진단의 의의와 활용에 대한 연구 (Study on the Significance and Application of the whole Body-form Diagnosis)

  • 김경철;신순식;이용태
    • 동의생리병리학회지
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    • 제16권5호
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    • pp.873-880
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    • 2002
  • We study on the significance and application of the whole body form diagnosis. The results were as follows; The general form diagnosis is the method to observe the individual physiology and pathology. The phase of thinking, the current and activity of KI, the pattern of general form diagnosis have organic relations with the symptoms. The general form diagnosis is made up the principle of the imaging phase, therefore it must make synthetic union the differentiation of syndromes. The general form diagnosis of NAE GYEONG shows the typical phases and it is divided with the sight of YIN YANG and Five-Element. The general form diagnosis of SEOP GAE is practiced the theory of constitution's demonstration before the understanding of symptoms. Then JANG NAM tried the type of constitution's demonstration. The general form diagnosis of DONG MU becomes the diagnostic root of constitution's demonstration in four type constitution theory.

Arthrobacoer sp. L-3가 생성하는 Glucose Isomerase의 최적 생성조건 (Efficient Production of Glucose Isomerase from Atrhrobacter sp. L-3)

  • 이은숙
    • 동아시아식생활학회지
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    • 제7권1호
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    • pp.29-33
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    • 1997
  • The efficient production of glucose isomerase (G. I0.) produced form Arthrobacter sp. L-3 was studied. The optimum culture time of the enzyme was 40hr. The maximum enzyme activity was found at glucose concentration 1%. G. I. activity did not affect inoculum size. The glucose isomerase activity was strongly influenced by the addition of glucose.

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백삼의 페놀산 조성과 항산화 활성 (Phenolic acid composition and antioxidative activity of white ginseng (Panax ginseng, C. A. Meyer))

  • 최창숙;김경임;홍희도;최상윤;이영철;김경탁;노정해;김성수;김영찬
    • Journal of Ginseng Research
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    • 제30권1호
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    • pp.22-30
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    • 2006
  • 본 연구는 백삼으로부터 페놀산 획분을 얻은 후 페놀산 성분들의 조성 및 함량을 조사하고, 이 획분 들의 항산화 활성을 측정하기 위해 실시되었다. 추출수율 및 페놀성 물질의 함유량은 유리형 (FPA)획분이 가장 높은 것으로 나타났고, GC 분석 결과에서도 페놀산 함유량이 높은 것으로 조사되었다. 그러나, 3종류의 페놀산 획분이 갖는 항산화 활성을 실험한 결과 ferulic acid함량이 가장 높았던 결합형 (BPA)페놀산 획분의 라디칼 소거 활성이 유의적으로 높은 것으로 조사되었다. 단, SOD유사활성에서는 에스테르형 페놀산 획분의 활성이 높은 것으로 나타났는데, 이것은 에스테르형 (EPA) 획분에서만 검출된 caffeic acid에 의한 활성으로 생각된다. 백삼의 페놀산 획분이 홍삼이나 다른 천연물의 추출물에 비해 in vitro에서 활성이 낮은 것으로 나타나지만, 동물실험 이나 임상실험에서는 어떠한 결과를 초래할 수 있을 지는 검토해 봐야 할 필요성이 있으므로, 본 연구의 결과는 그동안 사포닌 연구에 비해 많이 이루어지지 않았던 페놀산에 대한 연구뿐만 아니라 후속적으로 이뤄져야 할 임상실험을 위한 기초 자료로써의 가치가 있을 것으로 사료된다.

Aspergillus nidulans 영양균사에서 효소전구체형 ChsC 활성의 검출 (Detection of Zymogenic ChsC Activity in Vegetative Hyphae of Aspergillus nidulans.)

  • 박범찬;박윤희;박희문
    • 미생물학회지
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    • 제40권2호
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    • pp.178-182
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    • 2004
  • Saccharomyces cerevisiae의 class I 키틴생합성 효소인 Chsl의 활성측정법을 적용하여 Aspergillus nidulans의 영양균사에서 class I 키틴생합성 효소인 ChsC의 활성측정을 시도하였다. 그 결과, A. nidulans의 class I 키틴생합성효소도 효모류의 경우와 마찬가지로 트립신 처리에 의하여 활성화되는 효소전구체형태(zymogenic form)로 존재함을 알 수 있었다. 그리고 A. nidulans 야생형의 class I 키틴 생합성 효소활성은 트립신 처리에 의하여 6배 가량 증가되었다. 반면, chsC 유전자가 파괴된 돌연변이주는 트립신 처리에 의하여 효소활성이 증가되지 아니하였을 뿐만 아니라, 효소활성의 수준도 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성과 거의 동일한 수준이었다. 따라서, 트립신을 처리하여 측정한A. nidulans 야생형의 class I 키틴생합성 효소활성 값에서 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성을 제외한 값이 A. nidulans 야생형의 ChsC 효소활성임을 알았다. 이러한 조건을 토대로 영양균사 생장과정 동안 ChsC의 효소활성을 측정한 결과, chsC 유전자의 발현양상과 유사하게 액체배양상태의 영양균사가 무성분화능을 획득하는 시기로 알려진 시간대에 효소의 활성이 증가하였다. 이러한 결과는 이미 보고된 바와 같이 chs 유전자가 A. nidulans의 영양균사 생장에 관여함을 시사하고 있다.