• 제목/요약/키워드: ion-binding

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사람 양수중 다종의 세포외성 포스포리파제의 $A_2$의 부분정제 및 특성 (Partial Purification and Characterization of Multiple Forms of Extracellular Phospholipase $A_2$ in Human Amniotic Fluid)

  • 전용주;백석환;이지혜;문태철;민병우;장현욱
    • 약학회지
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    • 제41권2호
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    • pp.212-219
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    • 1997
  • Multiple forms of extracellular phospholipase $A_2$ have been detected in human amniotic fluid (HAF). When HAF was subjected to heparin-Sepharose column chromatography, phospholipase $A_2$ activity was detected in both heparin-non binding and binding fraction. The activity of heparin-non binding fraction was further purified by sequential uses of column chromatographies on butyl-Toy-opearl 650M and DEAE-Sephacel. DEAE-Sephacel fraction contained three different phospholipase $A_2$ activities (Peak I, II, III). The molecular weight of DEAE-Sephacel fraction phospholipase $A_2$ determined by SDS-PAGE were about 52KDa (Peak I). Peak II, III required micromolar $Ca^{2+}$ ion for its maximum activity, but Peak I enzyme showed calcium independent phospholipase $A_2$ activity and showed broad range of pH (6.0~10.0) optimum. All these enzymes were not recognized by a monoclonal antibody raised against phospholipase $A_2$ from human synovial fluid. These results suggest that HAF might contain multiple forms of extracellular phospholipase $A_2$, which may neither belong to the 14KDa group II phospholipase $A_2$ family nor cytosolic phospholipase $A_2$.

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Structural Analyses of Zinc Finger Domains for Specific Interactions with DNA

  • Eom, Ki Seong;Cheong, Jin Sung;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
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    • 제26권12호
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    • pp.2019-2029
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    • 2016
  • Zinc finger proteins are among the most extensively applied metalloproteins in the field of biotechnology owing to their unique structural and functional aspects as transcriptional and translational regulators. The classical zinc fingers are the largest family of zinc proteins and they provide critical roles in physiological systems from prokaryotes to eukaryotes. Two cysteine and two histidine residues ($Cys_2His_2$) coordinate to the zinc ion for the structural functions to generate a ${\beta}{\beta}{\alpha}$ fold, and this secondary structure supports specific interactions with their binding partners, including DNA, RNA, lipids, proteins, and small molecules. In this account, the structural similarity and differences of well-known $Cys_2His_2$-type zinc fingers such as zinc interaction factor 268 (ZIF268), transcription factor IIIA (TFIIIA), GAGA, and Ros will be explained. These proteins perform their specific roles in species from archaea to eukaryotes and they show significant structural similarity; however, their aligned amino acids present low sequence homology. These zinc finger proteins have different numbers of domains for their structural roles to maintain biological progress through transcriptional regulations from exogenous stresses. The superimposed structures of these finger domains provide interesting details when these fingers are applied to specific gene binding and editing. The structural information in this study will aid in the selection of unique types of zinc finger applications in vivo and in vitro approaches, because biophysical backgrounds including complex structures and binding affinities aid in the protein design area.

이온교환된 X형 및 Y형 제올라이트의 O1S 전자에 대한 X선 광전자분광학적 해석 (XPS Studies for O1S Electrons of Ion Exchanged Zeolite X and Y)

  • 김명철;김종택
    • 공업화학
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    • 제3권2호
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    • pp.240-246
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    • 1992
  • X선 광전자분광법을 이용하여 교환된 제올라이트내 산소원자의 전자구조를 조사하였다. $Na^+$, $Fe^{2+}$$Fe^{3+}$로 교환된 X형 및 Y형 제올라이트에 대하여 얻어진 $O_{1S}$스펙트럼은 골격산소의 결합에너지를 밝히기 위하여 분해하였다. 각 스펙트럼에서는 2-3개의 분해띠들이 있었다. 분해된 각 띠들의 특성은 교환된 양이온에 따라서 $O_{1S}$ 전자의 결합에너지와 띠면적의 관점에서 고찰되었다. 분해된 띠들을 각각 골격내 가교산소에 의한 것(띠 1), 양이온에 결합된 산소에 의한 것(띠 2), 양이온에 배위된 물분자내 산소에 의한 것(띠 3)으로 해석되었다. $O_{1S}$스펙트럼 면적의 대부분을 차지하고 있는 띠 1은 제올라이트내 Al함량의 감소에 따라서 결합에너지상의 증가를 나타내었다.

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CA2 혼입 페이스트의 부식저항성에 관한 연구 (A Study on Corrosion Resistance of CA2-Mixed Paste)

  • 김재돈;장일영
    • 한국산업융합학회 논문집
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    • 제25권2_2호
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    • pp.289-297
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    • 2022
  • Deterioration in durability of structures due to the steel corrosion is difficult to determine whether or not corrosion is initiated and how much propagated, and moreover, repair and maintenance are not easy to deal with. Therefore, preventive treatments can be the best option to avoid the deterioration. Various methods for preventing corrosion of steel, such as electrochemical treatments, anti-corrosion agents and steel surface coatings, are being developed, but economic and environmental aspects make it difficult to apply them to in-situ field. Thus, the purpose of this study was to improve corrosion resistance by using CA-based clinker that are relatively simple and expected to be economically profitable Existing CA-based clinkers had problems such as flash setting and low strength development during the initial hydration process, but in order to solve this problem, CA clinker with low initial reactivity were used as binder in this study. The cement paste used in the experiments was replaced with CA2 clinker for 0%, 10%, 20%, and 30% in OPC. And the mixture used in the chloride binding test for the extraction of water-soluble chloride was intermixed with Cl- 0.5%, 1%, 2%, and 3% by weight of binder content. To evaluate characteristic of hydration heat evolution, calorimetry analysis was performed and simultaneously chloride binding capacity and acid neutralization capacity were carried out. The identification of hydration products with curing ages was verified by X-ray diffraction analysis. The free chloride extraction test showed that the chlorine ion holding ability improved in order OC 10 > OC 30 > OC 20 > OC 0 and the pH drop resistance test showed that the resistance capability in pH 12 was OC 0 > OA 10 > OA 20 > OA 30. The XRD analyses showed that AFm phase, which can affect the ability to hold chlorine ions, tended to increase when CA2 was mixed, and that in pH12 the content of calcium hydroxide (Ca(OH)2), which indicates pH-low resistance, decreased as CA2 was mixed

제조방법별 솔잎추출물의 항산화성 검토 (Antioxidants of Pine Needle Extracts According to Preparation Method)

  • 김수민;김은주;조영석;성삼경
    • 한국식품과학회지
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    • 제31권2호
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    • pp.527-534
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    • 1999
  • 솔잎의 제조방법별 열수추출물(HPNP, CPNP, FPN)의 항산화성을 검토하기 위하여 in vitro상에서 실험을 수행하였다. 각 추출물들의 지방산화 억제능을 측정한 결과, $Fe^{2+}$이온 첨가구에서는 HPNP와 FPN이 $Fe^{2+}$ 이온 binding능력이 우수하였으나, CPNP는 오히려 지방산화를 촉진하였고, $Fe^{3+}$이온 첨가구에서는 $Fe^{3+}$이온 binding능력은 없는 것으로 나타났다. 활성 산소종 존재 하에서 솔잎추출물의 첨가가 지방산화에 미치는 영향은 활성산소종보다 지방산화도가 오히려 높게 나타났으며, 가장 낮은 값을 나타낸 FPN도 $KO_2$보다 높게 나타나 포집능력은 없는 것으로 나타났다. 지방산화를 촉진하는 iron 함량은 ferrous iron과 total iron 모두 CPNP가 가장 높게 나타났고, 다음으로 HPNP, FPN순으로 나타났다. 항산화제인 ascorbic acid함량은 FPN, HPNP, CPNP 순으로 나타났다. 산화성 활성 free radical에 전자를 공여하여 산화를 억제시키는 척도인 전자공여능은 HPNP, CPNP, FPN순이었고, HPNP의 전자공여능이 가장 높은 것으로 나타났다. Speroxide dismutase (SOD) 유사활성을 측정한 결과 HPNP, CPNP, FPN순으로 나타나 SOD 유사활성능은 HPNP가 가장 우수하였다. 솔잎추출물을 각기 다른 pH에서 측정한 아질산염 분해능은 반응용액의 pH가 낮을수록 아질산염 분해능이 높게 나타났고 특히, pH 1.2에서 높은 분해능을 나타내었다.

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식물에 의한 납, 카드뮴 흡수 기작에 미치는 자유이온 및 유기산-중금속 복합체의 영향 (Effects of free metal ions and organo-metal complexes on the absorption of lead and cadmium by plants)

  • 이미나;서병환;김권래
    • Journal of Applied Biological Chemistry
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    • 제64권2호
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    • pp.159-164
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    • 2021
  • 토양 중 중금속은 자유이온, 유기산-중금속 복합체를 포함한 다양한 화학종으로 존재한다. 이 중 중금속 자유이온 비율은 식물체의 중금속 흡수와 밀접한 관련이 있는 것으로 알려져 있다. 본 연구는 중금속 자유이온과 유기산-중금속 복합체 존재 비율이 식물의 납, 카드뮴 흡수에 어떤 영향을 미치는지 알아보기 위하여 수행되었다. 이를 위해 순환식 양액재배 장치에서 28일간 키운 상추를 48시간 동안 납과 카드뮴 용액에 침지시키는 실험을 진행하였다. 침지 전 시트르산, 옥살산, 아세트산, 휴믹산을 이용해 납과 카드뮴 용액의 중금속 자유이온과 유기산-중금속 복합체의 비율이 100:0, 90:10, 70:30, 60:40이 되도록 조절하였다. 침지 후 상추 뿌리와 잎의 납, 카드뮴 농도를 분석한 결과, 용액 중 중금속 자유이온의 비율과 식물이 흡수한 납, 카드뮴 농도는 상관관계를 보이지 않았다. 반면에 처리한 유기산의 종류와 중금속 종류에 따른 차이가 더 큰 것으로 나타났다. 시트르산은 납 흡수는 증가시켰지만 카드뮴 흡수는 감소시켰다. 시트르산은 다른 유기산보다 상추의 납 흡수율을 높였는데 이는 시트르산이 분자량이 크기 때문에 중금속 이온과 복합체를 형성할 수 있는 용량이 더 크기 때문인 것으로 보인다. 그러므로 본 연구 결과는 식물에 의한 중금속 흡수가 단순히 자유이온의 양에 따라 달라지기 보다는 존재하는 유기산의 분자량에 의한 결합용량 차이가 중금속 종류에 따라 달리 작용하여 결정된다는 것을 보여준다.

Construction of Two Metal-ion Binding Sites to Improve the 3′-5′Exonuclease Activity of Taq DNA Polymerase

  • Park, Yong-Hyun;Kim, Jong-Moon;Choi, Hye-Ja;Kim, Seog-K.;Kim, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • 제8권5호
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    • pp.471-477
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    • 1998
  • Taq DNA polymerase from Thermus aquaticus is very useful in the polymerase chain reaction. Taq DNA polymerase is classified in the pol I family, represented by E. coli DNA polymerase I. The three-dimensional structural alignment of 3'-5'exonuclease domains from the pol I family DNA polymerases explains why Taq DNA polymerase does not carry out proofreading in polymerase chain reactions. Three sequence motifs, Exo I, II, and III, must exist to carry out 3'-5'exonuclease activity for proof- reading by a 3'-5'exonuclease reaction, but these are abolished in Taq DNA polymerase. The key catalytic module in 3'-5'exonuclease is two metal ions chelated by four active-site carboxylic amino acids. Taq DNA polymerase was mutagenized to construct the catalytic module in the active site. The circular dichroism technique supported the formation of the catalytic module, and the radioactive assay showed that the 3'-5'exonuclease activity doubled in the mutant Taq DNA polymerase.

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Stabilization of Compact Protein Structures by Macrocyclic Hosts Cucurbit[n]urils in the Gas Phase

  • Lee, Jong Wha;Park, Mi Hyun;Ju, Jeong Tae;Choi, Yun Seop;Hwang, Soo Min;Jung, Dong Jin;Kim, Hugh I.
    • Mass Spectrometry Letters
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    • 제7권1호
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    • pp.16-20
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    • 2016
  • Characterization of intact protein structures in the gas phase using electrospray ionization combined with ion mobility mass spectrometry has become an important tool of research. However, the biophysical properties that govern the structures of protein ions in the gas phase remain to be understood. Here, we investigated the impact of host-guest complexation of ubiquitin (Ubq) with macrocyclic host molecules, cucurbit[n]urils (CB[n]s, n = 6, 7), on its structure in the gas phase. We found that CB[n] complexation induces the formation of compact Ubq ions. Both CB[6] and CB[7] exhibited similar effects despite differences in their binding properties in solution. In addition, CB[n] attachment prevented Ubq from unfolding by collisional activation. Based on the experimental results, we suggest that CB[n]s prevent unfolding of Ubq during transfer to the gas phase to promote the formation of compact protein ions. Furthermore, interaction with positively charged residues per se is suggested to be the most important factor for the host-guest complexation effect.

Structure function relationships amongst the purple acid phosphatase family of binuclear metal-containing enzymes

  • Hamilton, Susan
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2003년도 제2차 연례학술대회 발표논문집
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    • pp.5-5
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    • 2003
  • The purple acid phosphatases comprise a family of binuclear metal-containing enzymes. The metal centre contains one ferric ion and one divalent metal ion. Spectroscopic studies of the monomeric, ${\sim}$36 kDa mammalian purple acid phosphatases reveal the presence of an Fe(III)Fe(II) centre in which the metals are weakly antiferromagnetically coupled, whereas the dimeric, ${\sim}$110 000 kDa plant enzymes contain either Fe(III)Zn(II) or Fe(III)Mn(II). The three dimensional structures of the red kidney bean and pig enzymes show very similar arrangements of the metal ligands but some significant differences beyond the immediate vicinity of the metals. In addition to the catalytic domain, the plant enzyme contains a second domain of unknown function. A search of sequence databases was undertaken using a sequence pattern which includes the conserved metal-binding residues in the plant and animal enzymes. The search revealed the presence in plants of a 'mammalian-type' low molecular weight purple acid phosphatase, a high molecular weight form in some fungi, and a homologue in some bacteria. The catalytic mechanism of the enzyme has been investigated with a view to understanding the marked difference in specificity between the Fe-Mn sweet potato enzyme, which exhibits highly efficient catalysis towards both activated and unactivated phosphate esters, and other PAPs, which hydrolyse only activated esters. Comparison of the active site structures of the enzymes reveal some interesting differences between them which may account for the difference. The implications fur understanding the physiological functions of the enzymes will be discussed.

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Alkali Metal Ion Sensing in Aqueous Media by a Dibenzo-16-crown-5 Chromoionophore

  • Hayashita, Takashi;Kenji, Kunogi;Takagi, Makoto;Lee, Jong Chan;Bartsch, Richard A.
    • 분석과학
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    • 제8권4호
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    • pp.793-798
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    • 1995
  • A new chromoionophore sym-(decyl)(2-hydroxy-5-nitrobenzyloxy)-dibenzo-16-crown-5 (1) has been synthesized for $Na^+$ photometry in aqueous media. Apparent $pK_a$ values of 1 in the presence of 0.10 M LiCl, NaCl, and KCl were measured by spectrophotometry in 50% 1,4-dioxane-50% water (v/v) and compared with the $pK_a$ of 8.68 in the presence of 0.10 M TMACl. A significant $pK_a$ shift to a lower pH was only observed for $Na^+$ (${\Delta}pH=1.31$) due to selective binding of 1 with $Na^+$. Based upon this $pK_a$ shift, chromoionophore 1 was found to selectively respond to $Na^+$ with a detection limit of $10^{-3}M$ and no interference from $K^+$ up to 0.05 M for detection of 10.0 mM $Na^+$ in 50% 1,4-dioxane-50% water (v/v).

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