• 제목/요약/키워드: Cofactor

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HQNO-sensitive NADH:Quinone Oxidoreductase of Bacillus cereus KCTC 3674

  • Kang, Ji-Won;Kim, Young-Jae
    • BMB Reports
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    • 제40권1호
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    • pp.53-57
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    • 2007
  • The enzymatic properties of NADH:quinone oxidoreductase were examined in Triton X-100 extracts of Bacillus cereus membranes by using the artificial electron acceptors ubiquinone-1 and menadione. Membranes were prepared from B. cereus KCTC 3674 grown aerobically on a complex medium and oxidized with NADH exclusively, whereas deamino-NADH was determined to be poorly oxidized. The NADH oxidase activity was lost completely by solubilization of the membranes with Triton X-100. However, by using the artificial electron acceptors ubiquinone-1 and menadione, NADH oxidation could be observed. The activities of NADH:ubiquinone-1 and NADH:menadione oxidoreductase were enhanced approximately 8-fold and 4-fold, respectively, from the Triton X-100 extracted membranes. The maximum activity of FAD-dependent NADH:ubiquinone-1 oxidoreductase was obtained at about pH 6.0 in the presence of 0.1M NaCl, while the maximum activity of FAD-dependent NADH:menadione oxidoreductase was obtained at about pH 8.0 in the presence of 0.1M NaCl. The activities of the NADH:ubiquinone-1 and NADH:menadione oxidoreductase were very resistant to such respiratory chain inhibitors as rotenone, capsaicin, and $AgNO_3$, whereas these activities were sensitive to 2-heptyl-4-hydroxyquinoline-N-oxide (HQNO). Based on these results, we suggest that the aerobic respiratory chain-linked NADH oxidase system of B. cereus KCTC 3674 possesses an HQNO-sensitive NADH:quinone oxidoreductase that lacks an energy coupling site containing FAD as a cofactor.

Unusual malignant neoplasms occurring around dental implants: A report of 2 cases

  • Oh, Song Hee;Kang, Ju Hee;Seo, Yu-Kyeong;Lee, Sae Rom;Choi, Yong-Suk;Hwang, Eui-Hwan
    • Imaging Science in Dentistry
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    • 제48권1호
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    • pp.59-65
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    • 2018
  • Osseointegrated implants are now commonplace in contemporary dentistry. However, a number of complications can occur around dental implants, including peri-implantitis, maxillary sinusitis, osteomyelitis, and neoplasms. There have been several reports of a malignant neoplasm occurring adjacent to a dental implant. In this report, we describe 2 such cases. One case was that of a 75-year-old man with no previous history of malignant disease who developed a solitary plasmacytoma around a dental implant in the left posterior mandible, and the other was that of a 43-year-old man who was diagnosed with squamous cell carcinoma adjacent to a dental implant in the right posterior mandible. Our experiences with these 2 cases suggest the possibility of a relationship between implant treatment and an inflammatory cofactor that might increase the risk of development of a malignant neoplasm.

Interaction of Norfloxacin with Super-Coiled DNA

  • Hwangbo, Hyun-Jung;Lee, Young-Ae;Park, Jung-Hag;Lee, Yong-Rok;Kim, Jong-Moon;Yi, Seh-Yoon;Kim, Seog K.
    • Bulletin of the Korean Chemical Society
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    • 제24권5호
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    • pp.579-582
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    • 2003
  • Norfloxacin, that inhibits the action of topoisomerase Ⅱ, binds to wide variety of DNA. The binding mode of this drug to double- and super-coiled DNA (ds- and scDNA) is compared in this study by various spectroscopic methods, including absorption, fluorescence, and circular dichroism(CD) spectroscopy. Hypochromism in the absorption band, negative and positive induced CD bands (respectively in 240-260 nm and 270-300 nm region) are apparent for the norfloxacin that bound to both the dsDNA and scDNA. A decrease in fluorescence is also noticed in the presence of both DNAs. Since the spectroscopic characteristics are the same for both complexes, it is imperative that the binding mode of the norfloxacin is similar in ds- and scDNA. In the presence of $Mg^{2+}$, which is a cofactor in the topoisomerase Ⅱ action, the fluorescence intensity of the scDNA-norfloxacin complex increased and the resulting fluorescence intensity and shape was identical to that in the absence of scDNA. Therefore, the addition of an excess amount of $Mg^{2+}$ may result in the extrusion of norfloxacin from scDNA.

분자공학 기반의 광학활성 에폭사이드 제조용 epoxide hydrolase 생촉매 개발 (Molecular Engineering of Epoxide Hydrolases for Production of Enantiopure Epoxides)

  • 김희숙;이은열
    • 생명과학회지
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    • 제16권1호
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    • pp.168-174
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    • 2006
  • 광학활성 에폭사이드는 다양한 반응성으로 인하여 고부가가치 광학활성 의약품 및 농약 합성용 중간체로 널리 이용되고 있다. 광학활성 에폭사이드는 에폭사이드 가수분해효소 (epoxide hydrolase, EH)를 이용하여 저가의 라세믹 기질에 대한 입체선택적 가수분해 반응을 통해 제조할 수 있으며, EH는 유도과정 없이 발현되고 보조인자가 필요 없으며 비교적 효소 안정성도 높아 상업적으로 유용한 효소이다. EH에 대한 생화학 및 분자생물학 관련 최근 연구 결과를 바탕으로 촉매 활성 증대 및 기질 선택성을 변경시킨 tailer-made형 EH 생촉매 개발이 가능할 것이며, 실규모의 비대칭 광학분할 생물공정 시스템 개발을 통해 EH에 의한 동력학적 가수분해반응을 이용한 광학활성 에폭사이드 생산기술의 상업화가 가능할 것으로 기대된다.

Biochemical and molecular characterization of a tetrachloroethylene (PCE) dechlorinating Clostridium bifermentans DPH-1

  • Chang, Young-Cheol;Toyama, Tadashi;Kikuchi, Shintaro
    • 환경위생공학
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    • 제23권2호
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    • pp.1-18
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    • 2008
  • The tetrachloroethylene (PCE) dehalogenase of Clostridium bifermentans DPH-1 (a halorespiring organism) was purified, cloned, and sequenced. This enzyme is a homodimer with a molecular mass of ca. 70 kDa and exhibits dehalogenation of dichloroethylene isomers along with PCE and trichloroethylene (TCE). Broad range of substrate specificity for chlorinated aliphatic compounds (PCE, TCE, cis-1,2-dichloroethylene, trans-1,2-dichloroethylene, 1,1-dichloroethylene, 1,2-dichloropropene, and 1,1,2-trichloroethane) for this enzyme was also observed. A mixture of propyl iodide and titanium citrate caused a light-reversible inhibition of enzymatic activity suggesting the involvement of a corrinoid cofactor. A partial sequence (81 bp) of the encoding gene for PCE dehalogenase was amplified and sequenced with degenerateprimers designed from the N-terminal sequence (27 amino acid residues). Southern analysis of C. bifermentans genomic DNA using the polymerase chain reaction product as a probe revealed restriction fragment bands. A 5.0 kb ClaI fragment, harboring the relevant gene (designated pceC) was cloned (pDEHAL5) and the complete nucleotide sequence of pceC was determined. The gene showed homology mainly with microbial membrane proteins and no homology with any known dehalogenase, suggesting a distinct PCE dehalogenase. So, C. bifermentans could play some important role in the initial breakdown of PCE and other chlorinated aliphatic compounds in sites contaminated with mixtures of halogenated substances.

Dihydropyrrolo[3,4-f]quinazoline 엽산길항제의 합성 및 In Vitro 항암활성 (Synthesis of Dihydropyrrole[3,4-f]quinazoline Antifolates and Their Antitumor Activity In Vitro)

  • 백두종
    • 약학회지
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    • 제50권4호
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    • pp.278-286
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    • 2006
  • Classical dihydropyrrole[3,4-f]quinazoline antifolates 7,8 and 9, in which the tricyclic ring is structurally similar to the pteridine ring of $CH_2-THF(1)$, the cofactor of thymidylate synthase (TS), were synthesized, and their in vitro antitumor activity was evaluated by measuring the cell growth inhibitory activity against cancer cell lines. The target compounds were cytotoxic against CCRF-CEM, human T-cell acute lymphoblastic leukemia, with the cell growth inhibitory activity $(IC_{50})$ of $0.8{\sim}8.3\;{\mu}M$. Among the three compounds, 3-amino analog 7 was 10- and 3.5-fold more cytotoxic compared to the 3-methyl analogs 8 and 9, and its cytotoxicity was similar to that of the reference compound with the $IC_{50}$ value of $0.83\;{\mu}M$. This result was supposed as the consequence of the fact that dihydropyrroloquinazolinone ring with amino group was able to bind well in the active site of TS. In the case of 3-methyl analogs, analog 9, which has two-carbon bridge between the dihydropyrroloquinazolinone ring and benzoyl-L-glutamic acid, was 3-times more potent in cytotoxicity than analog 8 which has one-carbon bridge, and this result indicates that the distance and conformational orientation of the benzoyl-L-glutamic acid moiety with respect to the tricyclic ring may also be a crucial determinant of cell growth inhibitory activity.

Regulatory Mechanism of L-Alanine Dehydrogenase from Bacillus subtilis

  • 김수자;김유진;서미란;전봉숙
    • Bulletin of the Korean Chemical Society
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    • 제21권12호
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    • pp.1217-1221
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    • 2000
  • L-alanine dehydrogenase from Bacillus subtilis exhibits allosteric kinetic properties in the presence of $ZN^{2+}$. $ZN^{2+}$ induces the binding of substrate (L-alanine) to be cooperative at pH 8.0. The effect of pH variation between pH 7.0 and pH 10.0 on the inhibition by $ZN^{2+}$ correlates with the pH effect on the $K_m$ values for L-alanine within these pH range indicating that $ZN^{2+}$ and substrate compete for the same site. No such cooperativity is induced by $ZN^{2+}$ when the reaction is carried out at pH 10. At this higher pH, $ZN^{2+}$ binds with the enzyme with lower affinity and noncompetitive with respect to L-alanine. Inhibition of L-alanine dehydrogenase by $ZN^{2+}$ depends on the ionic strength. Increase in KCI concentration reduced the inhibition, but allosteric property in $ZN^{2+}$ binding is conserved. A model for the regulatory mechanism of L-alanine dehydrogenase as a noncooperative substrate-cooperative cofactor allosteric enzyme, which is compatible in both concerted and the sequential allosteric mechanism, is proposed.

Regulatory Role of Zinc in Immune Cell Signaling

  • Kim, Bonah;Lee, Won-Woo
    • Molecules and Cells
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    • 제44권5호
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    • pp.335-341
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    • 2021
  • Zinc is an essential micronutrient with crucial roles in multiple facets of biological processes. Dysregulated zinc homeostasis impairs overall immune function and resultantly increases susceptibility to infection. Clinically, zinc supplementation is practiced for treatment of several infectious diseases, such as diarrhea and malaria. Recent focus on zinc as a beneficial element for immune system support has resulted in investigation of the immunomodulatory roles of zinc in a variety of immune cells. Besides its classical role as a cofactor that regulates the structural function of thousands of proteins, accumulating evidence suggests that zinc also acts, in a manner similar to calcium, as an ionic regulator of immune responses via participation as an intracellular messenger in signaling pathways. In this review, we focus on the role of zinc as a signaling molecule in major pathways such as those downstream of Toll-like receptors-, T cell receptor-, and cytokine-mediated signal transduction that regulate the activity and function of monocytes/macrophages and T cells, principal players in the innate and adaptive immune systems.

A Collaborative Study to Establish a Korea National Biological Standard for Antithrombin Concentrate

  • Kang, Hye-Na;Lee, Sung-Han;Kim, Soon-Nam;Hong, Choong-Man;Lee, Ki-Hong;Oh, Ho-Jung;Yoo, Si-Hyung;Shin, In-Soo;Choi, Seung-Eun;Lee, Seok-Ho;Gray, Elaine;Okada, Yoshiaki;Hong, Seung-Hwa
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.272.1-272.1
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    • 2002
  • We have carried out collaborative study to evaluate a preparation of antithrombin concentrate whether or not it was suitable to serve as the candidate for a Korea National Biological Standard. Six laboratories. including three manufacturers and three National Control Laboratories. participated in this study. The potency of this candidate preparation was determined using the heparin cofactor chromogenic method. (omitted)

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Inactivation of Brain GABA transaminase by p$^1$, p$^2$-Bis(5′-pyridoxal) diphosphate

  • Jang, S.H.;Lee, B.R.;J.W. Hong;Park, K.W.;Yoo, B.K.;Cho, S.W.;Park, S.Y.
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1995년도 춘계학술대회
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    • pp.74-74
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    • 1995
  • GABA transaminase is inactivated by preincubation with p$^1$, p$^2$-bis(5'-pyridoxal) diphosphate at pH 7.0. The inactivation under pseudo-first order conditions proceeds at a slow rate (K$\_$obs/=0.035 min$\^$-1/). The degree of labeling of the enzyme by p$^1$, p$^2$-bis(5'-pyridoxal) diphosphate was determined by absorption spectroscopy, The blocking of 2 lysyl residues/dimer is needed for inactivation of the transaminase. The time course of the reaction is significantly affected by the substrate ${\alpha}$-ketoglutarate, which afforded complete protection against the loss of the catalytic activity. Whereas cofator pyridoxal phosphate failed to prevent the inactivation of the enzyme. Therefore, it is postulated that binding of ${\alpha}$-ketoglutarate tn lysyl residues is the major factor contributing to stabilization of the catalytic site and bifuctional reagent p$^1$, p$^2$bis(5'-pyridoxal) diphosphate blocks lysyl residues other than those involved in the binding of the cofactor.

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