• Biomolecules & Therapeutics
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• 제22권5호
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• pp.414-419
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• 2014

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that regulate cell-matrix composition and are also involved in processing various bioactive molecules such as cell-surface receptors, chemokines, and cytokines. Our group recently reported that MMP-3, -8, and -9 are upregulated during microglial activation and play a role as proinflammatory mediators (Lee et al., 2010, 2014). In particular, we demonstrated that MMP-8 has tumor necrosis factor alpha (TNF-${\alpha}$)-converting enzyme (TACE) activity by cleaving the prodomain of TNF-${\alpha}$ and that inhibition of MMP-8 inhibits TACE activity. The present study was undertaken to compare the effect of MMP-8 inhibitor (M8I) with those of inhibitors of other MMPs, such as MMP-3 (NNGH) or MMP-9 (M9I), in their regulation of TNF-${\alpha}$ activity. We found that the MMP inhibitors suppressed TNF-${\alpha}$ secretion from lipopolysaccharide (LPS)-stimulated BV2 microglial cells in an order of efficacy: M8I>NNGH>M9I. In addition, MMP inhibitors suppressed the activity of recombinant TACE protein in the same efficacy order as that of TNF-${\alpha}$ inhibition (M8I>NNGH>M9I), proving a direct correlation between TACE activity and TNF-${\alpha}$ secretion. A subsequent pro-TNF-${\alpha}$ cleavage assay revealed that both MMP-3 and MMP-9 cleave a prodomain of TNF-${\alpha}$, suggesting that MMP-3 and MMP-9 also have TACE activity. However, the number and position of cleavage sites varied between MMP-3, -8, and -9. Collectively, the concurrent inhibition of MMP and TACE by NNGH, M8I, or M9I may contribute to their strong anti-inflammatory and neuroprotective effects.

• 한국식품영양과학회지
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• 제23권3호
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• pp.490-495
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• 1994

Acid phosphatase (EC3.1.3.2) from carrots was partially purified by ammonium sulfate fractionation (30%-80%), Sephacryl S-200 gel filtration, cm-Sepharose CL-6B and DEAE -Sephacel ion exchange chromatography. The optimum ph and temperature of acid phosphatase from carrots were pH 5.5 and 55$^{\circ}C$, respectively. The enzyme was most stable at ph 6.0 and relatively unstable below pH 4.0 . The activation energy of the enayme was determined to be 10.6kcal/mole. The enzyme utilized p-nitrophenyl phosphate as a substrate among tested possible substrates, whereas it hydrolyzed 5' -IMP and 5'-GMP poorly. The Michaelis -Menten constant(Km) of the enzyme with p-nitrophenyl phosphate as a substrate was identified as 0.55mM. Amongtested metal ions and inhibitors, Al+++ Zn++, Cu++ , fluoride, metavanadate and molybdate ions inhibited the enzyme activity drastically.

• Applied Biological Chemistry
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• 제41권8호
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• pp.608-610
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• 1998

To investigate the inhibitory activity of 2-hydroxy-3-amino-4-phenylbutyrate-harboring aminopeptidase N inhibitors, p-nitro-AHPA-peptide derivatives (1 and 2) and an AHPA-peptide derivative (3) were synthesized by chain elongation from C-terminal end using DCC/HOBt as a coupling reagent. The peptides $1{\sim}3$ exerted strong inhibitory activities against aminopeptidase N with $IC_{50}$ values of 1.8, 7.3 and $24.0\;{\mu}g/ml$, respectively, and cytotoxicity on cancer cell lines in vitro.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.638-639
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• 2000

Quinone reductase was purified to electrophoretic homogeneity from bovine liver by using ammonium sulfate fractionation, ion-exchange chromatography, and gel filtration chromatography. The enzyme utilized either NADH or NADPH as the electron donor. The optimum pH of the enzyme was pH 8.5, and the activity of the enzyme was greatly inhibited by $Cu^{2+}$ and $Hg^{2+}$ ions, dicumarol and cibacron blue 3GA. The enzyme catalyzed the reduction of several quinones and other artificial electron acceptors. Furthermore, the enzyme catalyzed NAD(P)H-dependent reduction of azobenzene or 4-nitroso-N,N-dimethylaniline. The apparent $K_m$ for 1,4-benzoquinone, azobenzene, and 4-nitroso-N,N-dimethylaniline was 1.64mM, 0.524mM and 0.225mM, respectively. The reduction of azobenzene or 4-nitroso-N,N-dimethylaniline by quinone reductase was strongly inhibited by dicumarol or cibacron blue 3GA, potent inhibitors of quinone reductase.

• 정신신체의학
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• 제19권2호
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• pp.57-65
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• 2011

많은 심혈관질환약물과 향정신성약물 간에 다양한 약물상호작용이 존재하며 이러한 약물들의 대부분이 시트크롬(cytochrome, CYP)450 효소의 기질, 억제제, 유도제로 작용하면서 약물상호작용이 일어나게 된다. 주로 CYP2D6와 CYP3A4를 억제하는 향정신성약물로 인해 같이 투여되는 심혈관질환약물의 효과가 변할 수 있고 부작용까지 나타날 수 있다. 이런 상황을 고려하고 반대의 경우도 포함하여 흔히 처방되는 두 종류의 약물을 병용 투여하는 경우 고려해야 할 부분에 대해서 심혈관질환약물 분류에 따라 논하였다. 대부분의 베타차단제는 CYP2D6의 대사에 의존하므로 이 대사를 억제하는 bupropion, chlorpromazine, haloperidol, SSRIs, quinidine 등을 사용했을 때 베타차단제의 독성이 나타날 수 있다. 앤지오텐신 관련 약물과 이뇨제가 lithium의 농도를 변화시키는 점도 고려하여야 한다. 칼슘통로차단제 및 콜레스테롤강하제를 CYP3A4의 강력한 억제제인 amiodarone, diltiazem, fluvoxamine, nefazodone, verapamil 등과 함께 사용하였을 때 약물 상호작용에 따른 부작용에 유의하여야 한다. 항부정맥제를 복용하는 환자에서 QT 간격 증가를 야기하는 약물이나 관련 CYP450 효소를 억제하는 약물을 동시에 투여하는 것은 삼가거나 적극적인 관찰이 필요하다. Digoxin과 warfarin이 병용 투여되는 향정신성약물로 인해 혈중 농도가 변하는 것도 임상적으로 중요하다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1999년도 춘계학술대회
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• pp.5-8
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• 1999

여성의 breast cancer의 1/3 의 경우가 hormone-dependent에 의한 것이다. 따라서 그 치료의 접근 방법으로써 estrogen receptor에 작용하거나, receptor-mediated gene trenscription을 저해하는 antiestrogen을 사용하는 것이 있다. Estrogen은 microsomal cytochrome P-450 enzyme complex system에 의해 androgen으로부터 생합성되는 hormone이며, estrogen product는 steroid product의 biosynthetic sequence의 마지막 단계에서 생산되고, aromatase는 이에 관여하는 enzyme으로써, aromatase를 선택적으로 저해하는 경우에 다른 steroid의 생산에 영향을 미치지 않고 estrogen 생산을 감소 시킬 수 있다. 이러한 관점에서, aromatase를 선택적으로 저해하는 것은 hormone-dependent breast cancer를 완화시킬 수 있는 가능성을 가진 cancer chernopreventive agents의 새로운 방법이라 할 수 있다.

• Bulletin of the Korean Chemical Society
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• 제27권10호
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• pp.1567-1571
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• 2006

Synthetic color additives approved for general food use are sixteen in European Union, seven in U. S. A. and twelve in Japan. Twelve food dyes were examined for their inhibitory potency against human protein tyrosine phosphatases (PTPases). Half of the food colorants inhibited PTPases significantly and three of them were potent inhibitors with low micromolar IC50 values. Also examined were the synthetic dyes structurally similar but not allowed in food. Some of them were potent inhibitors of PTPases. Considering the importance of PTPases in cellular signal transduction, inhibition of PTPases by food colorants might cause harmful effects in human health.

• Biomolecules & Therapeutics
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• 제3권1호
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• pp.34-37
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• 1995

Non-steroidal anti-inflammatory drugs (NSAIDs) have been known as inhibitors of the folate-requiring enzymes. In the present work, we have expanded on these observations and have investigated the inhibitory effects of NSAIDs on Lactobacillus casei thymidylate synthase expressed in E. coli. NSAIDs including sulphasalizine, salicylic acid, indomethacin and mefenamic acid were found to be competitive inhibitors with respect to folate of Lactobacillus casei thymidylate synthase. In contrast, aspirin and the antipyretic-analgesic drugs acetaminophen and antipyrine were weak inhibitors of the enzyme. Structure-activity correlation suggests that an aromatic ring with a side chain containing a carboxylic acid is a requirement for competitive inhibition of the thymidylate synthase. The results are consistent with the hypothesis that the antifolate activity of NSAIDs, and hence cytostatic consequences, are important factors in producing anti-inflammatory activity and aspirin exerts its anti-inflammatory effects after its conversion into salicylic acid, which possesses greater antifolate activity than its parent compound.

• 생명과학회지
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• 제20권6호
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• pp.838-844
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• 2010

토양에서 생육하는 Streptomyces corcohrussi의 혈전용해효소가 DEAE-Sephadex A-50 그리고 Sephadex G-50 젤 여과를 이용한 크로마토그라피 방법에 의해 순수분리 되었다. SDS-PAGE 분석결과, 분리된 효소는 단일 단백질이고, 그 분자량은 약 34 kDa 이라는 것을 알 수 있었다. 순수분리된 효소의 혈전용해활성은 plasminogen-rich fibrin plate에서 0.8 U/ml 이었으나, plasminogen-free fibrin plate 에서의 그 효소활성은 0.36 U/ml 이하이었다. 이러한 결과로, 순수 분리된 효소가 plasminogen activator 로 작용한다는 것을 알 수 있었다. 단백질 저해제인 $\varepsilon$-ACA, t-AMCHA 와 mercuric chloride의 존재시에 그 혈전용해활성은 24% 이하이었는데, 이러한 결과는 이들 plasmin 저해제 그리고(혹은) fibrinogen을 fibrin으로 전환시키는 과정과 관련된 fibrinogen 저해제에 의해 이 효소가 조절될 수 있음을 나타낼 수 있다. 한편으로, 중금속 이온인 $Zn^{2+}$은 그 활성을 58% 감소시켰다. 순수 분리된 효소의 최적 온도는 약 $50^{\circ}C$ 이었고, 그 효소활성의 92% 이상은 pH 5.0과 8.0 사이에서 유지되었다. 그러므로, 이러한 결과들은 하나의 강력한 혈전용해효소를 제공해서, S. corcohrussi 유래 새로운 혈전용해제의 개발에 기여하도록 한다.

• Fisheries and Aquatic Sciences
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• 제1권2호
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• pp.209-215
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• 1998

Trypsin-like enzyme was purified from shrimp hepatopancreas through Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Superdex 75 gel chromatography. Purity of trypsin-like enzyme was increased 69-fold with $44\%$ yield. The enzyme consisted of a single polypeptide chain with a molecular weight (M.W.) of 32 kDa judged by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was completely inactivated by serine enzyme inhibitors such as soybean trypsin inhibitor (SBTI), tosyl-L­lysine chloromethyl ketone (TLCK), and leupeptin. However, the enzyme was not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) which is a chymotrypsin specific inhibitor. The enzyme had no activity against benzoyl-tyrosine ethyl ester (BTEE) which is a chymotrypsin specific substrate. The enzyme showed high activity on the carboxyl terminal of Phe, Tyr. Glu, Arg, and Asp. However. no activity was detected against the carboxyl terminal of Pro, Trp, Cys, Gly, Val, and Ala.