• 제목/요약/키워드: Metal-binding protein

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무작위 액중 상호 작용에 의한 단백질칩의 개발 (Development of Protein Chip by Random Fluidic Self-Assembly Interaction)

  • 최용성;권영수;박대희
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2003년도 추계학술대회 논문집 전기물성,응용부문
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    • pp.303-305
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    • 2003
  • In this paper, we have been proposed a new method of multichannel biosensor using random fluidic self-assembly. A metal particle and an array was fabricated. Biomaterials were immobilized on the metal particle. The array and the particles were mixed in a buffer solution, and were arranged by self-assembly. A quarter of total Ni dots were covered by the particles. The binding direction of the particles was controllable, and condition of particles was almost with Au surface on top. The particles were successfully arranged on the array. The biomaterial activities were detected by chemiluminescence.

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자성체를 이용한 단백질칩 마이크로어레이의 개발 (Development of Protein Chip Microarray Using a Magnetic)

  • 최용성;이경섭;박대희
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2005년도 제36회 하계학술대회 논문집 C
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    • pp.2398-2400
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    • 2005
  • In this paper, we have been described a new constructing method of multichannel biosensor using self-assembly by magnetic force interaction. A metal particle and an array was fabricated by photolithographic. Biomaterials were immobilized on the metal particle. The array and the particles were mixed in a buffer solution, and were arranged by magnetic force interaction and self-assembly. A quarter of total Ni dots were covered by the particles. The binding direction of the particles was controllable, and condition of particles was almost with Au surface on top. The particles were successfully arranged on the array. The biomaterial activities were detected by chemiluminescence.

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자기력에 의한 단백질칩 (Protein Chip by Magnetic Force)

  • 최용성;문종대;이경섭
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2006년도 제37회 하계학술대회 논문집 C
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    • pp.1317-1318
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    • 2006
  • In this paper, we have been described a new constructing method of multichannel biosensor using self-assembly by magnetic force interaction. A metal particle and an array was fabricated by photolithographic. Biomaterials were immobilized on the metal particle. The array and the particles were mixed in a buffer solution, and were arranged by magnetic force interaction and self-assembly. A quarter of total Ni dots were covered by the particles. The binding direction of the particles was controllable, and condition of particles was almost with Au surface on top. The particles were successfully arranged on the array. The biomaterial activities were detected by chemiluminescence.

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자기력에 의한 단백질칩 (Protein Chip Using Magnetic Force)

  • 최용성;문종대;이경섭
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2006년도 하계학술대회 논문집 Vol.7
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    • pp.386-387
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    • 2006
  • This research describes a new constructing method of multifunctional biosensor using many kinds of biomaterials. A metal particle and an array was fabricated by photolithographic. Biomaterials were immobilized on the metal particle. The array and the particles were mixed in a buffer solution, and were arranged by magnetic force interaction and self-assembly. A quarter of total Ni dots were covered by the particles. The binding direction of the particles was controllable, and condition of particles was almost with Au surface on top. The particles were successfully arranged on the array. The biomaterial activities were detected by chemiluminescence and electrochemical methods.

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금속 친화성 액 이상분계 시스템에서 단백질의 분배 향상 (Enhanced Partitioning of Proteins in Metal-Affinity Aqueous Two-Phase Systems)

  • 정봉현;박영훈
    • 한국미생물·생명공학회지
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    • 제21권4호
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    • pp.336-341
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    • 1993
  • In metal-affinify aqueous two-phase systems, protein partitioning is affected by a variety of parameters such as pH, the number of surface-accessible histidines, and the amount and partition coefficient of metallated polythylene glyco(PEG) ligand. To enhance partitioning of proteins with surface-accessible histidines, we have synthesized and used a (Cu(II)-ininodiacetic acid)$_2$-PEG20,000($Cu(II)_2IDA_2$-PEG20,000) as well as Cu(II)IDA-PEG5,000 as an affinity ligand. The partition coefficient of $Cu(II)_2-IDA_2$-PEG20,000 in a PEG5,000/dextran two-phase system was 30.1, which corresponded to a 3.8-fold increase over that of Cu(II)IDA-PEG5,000. The partitioning experiments were performed on four proteins, horse cytochrome c, S. cerevisiae cytochrome c, horse myoglobin, and sheep myoglobin. Partitioning of proteins which convey surface-accessible histidines was enhanced dramatically by the addition of $Cu(II)_2IDA_2$-PEG20,000 ligand. These results demonstrate that enhanced partitioning of metal-binding proteins in an aqueous two -phase system can by achieved by using an appropriate metallated PEG ligand.

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Retrospective analyses of the bottleneck in purification of eukaryotic proteins from Escherichia coli as affected by molecular weight, cysteine content and isoelectric point

  • Jeon, Won-Bae
    • BMB Reports
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    • 제43권5호
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    • pp.319-324
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    • 2010
  • Experimental bioinformatics data obtained from an E. coli cell-based eukaryotic protein purification experiment were analyzed in order to identify any bottleneck as well as the factors affecting the target purification. All targets were expressed as His-tagged maltose-binding protein (MBP) fusion constructs and were initially purified by immobilized metal affinity chromatography (IMAC). The targets were subsequently separated from the His-tagged MBP through TEV protease cleavage followed by a second IMAC isolation. Of the 743 total purification trials, 342 yielded more than 3 mg of target proteins for structural studies. The major reason for failure of target purification was poor TEV proteolysis. The overall success rate for target purification decreased linearly as cysteine content or isoelectric point (pI) of the target increased. This pattern of pI versus overall success rate strongly suggests that pI should be incorporated into target scoring criteria with a threshold value.

Structural Analysis of [Cu(II)-amyloidogenic peptide] Complexes

  • Cha, Eugene;Seo, Jae-Hong;Kim, Ho-Tae
    • Mass Spectrometry Letters
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    • 제9권1호
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    • pp.17-23
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    • 2018
  • Studies on the interactions of amyloidogenic proteins with trace metals, such as copper, have indicated that the metal ions perform a critical function in the early oligomerization process. Herein, we investigate the effects of Cu(II) ions on the active sequence regions of amyloidogenic proteins using electrospray ionization mass spectrometry (ESI-MS) and collision induced dissociation tandem MS (CID-MS/MS). We chose three amyloidogenic peptides NNQQNY, LYQLEN, and VQIVYK from yeast prion like protein Sup35, insulin chain A, and tau protein, respectively. [Cu-peptide] complexes for all three peptides were observed in the mass spectra. The mass spectra also show that increasing Cu(II) concentrations decrease the population of existing peptide oligomers. The tandem mass spectrum of NNQQNY shows preferential binding for the N-terminal region. All three peptides are likely to appear to be in a Cu-monomer-monomer (Cu-M-M) structure instead of a monomer-Cu-monomer (M-Cu-M) structure.

Two-component System 기반 재조합균을 이용한 인공폐수에서의 중금속 인지 및 제거 (Heavy Metal Detection and Removal in Artificial Wastewater Using Two-Component System Based Recombinant Bacteria)

  • ;홍순호;유익근
    • 미생물학회지
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    • 제48권3호
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    • pp.187-191
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    • 2012
  • Two component system (TCS)인 ZraS/R 및 CusS/R의 zraP와 cusC 유전자의 프로모터에 의해 green fluorescent protein (GFP)이 발현되도록 제작된 박테리아 바이오센서의 성능을 인공폐수에서 평가하였다. 제작된 박테리아 바이오센서는 실제 폐수를 모사한 인공폐수에서도 시료 중의 $Zn^{2+}$$Cu^{2+}$를 인지하여 GFP를 효율적으로 발현시키는 것을 확인할 수 있었다. 두 번째는 세포 표면에 금속 친화성 펩타이드가 표현되도록 제작된 구리 및 아연 제거 박테리아를 인공폐수 조건에서 평가하였다. 제작된 박테리아는 각각 ZraS/R 및 CusS/R TCS에 의해 주변의 $Zn^{2+}$$Cu^{2+}$를 인지하여 세포 표면에 OmpC-ZBP와 OmpC-CBP 융합 단백질을 발현시키는 시스템이다. 실험을 통해 표면에 금속 흡착 펩타이드가 발현된 재조합 균은 인공폐수 조건에서도 효과적으로 구리 및 아연을 흡착시키는 것을 확인하였다. 따라서 본 연구에서 개발된 TCS 기반 재조합 균은 인공폐수 조건에서 중금속의 인지 및 제거 기능이 효과적으로 작동하는 것이 확인되었다.

말 비장 Ferritin에서 합성된 Al과 $UO_2$ 나노 입자의 전자현미경 연구 (Electron Microscopy of the Al and $UO_2$ Nanophase Particles Synthesized in Horse Spleen Ferritin)

  • 문향란;김경숙;이정후
    • Applied Microscopy
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    • 제29권3호
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    • pp.323-329
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    • 1999
  • 본 연구에서는 Al과 $UO_2$의 나노미터 크기의 무기물 입자를 합성하기 위하여 생체 시스템인 철단백질 ferritin을 이용하였다. Ferritin에서 합성된 Al과 $UO_2$의 무기물 입자를 TEM을 이용하여 관찰한 결과, 단백질 내부에 나노미터 크기의 구형인 미네랄(mineral) core를 확인하였다. 그 입자들에 대한 EDXA분석 결과 각각 Al과 $UO_2$로 구성된 미네랄임을 확인하였다. Ferritin을 이용하여 합성된 Al core는 이번 연구에서 처음으로 전자현미경으로 관찰되었다. 그리고 두 종류의 다른 금속 즉, Al/Fe및 $UO_2/Al$의 존재하에 ferritin core를 합성하여 TTM을 관찰한 결과, 역시 나노미터 크기의 구형인 전자밀도 core를 관찰하였고 EDXA분석 결과 구형인 core가 합성시킨 두 금속 원소의 미네랄로 구성되어 있음을 증명하였다. 그리하여 본 연구는 철단백질을 이용하여 철이 아닌 Al과 $UO_2$로 구성된 나노미터 크기의 무기물 입자를 합성할 수 있음을 증명하였다.

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금속 친화성 액 이상분계 시스템에서 Phsphoprotein분배 및 분배예측 (Phosphoprotein Partitioning in Metal-Affinity Aqueous Two-Phase Systems and Prediction of Partitioning Behavior)

  • 정봉현
    • KSBB Journal
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    • 제9권3호
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    • pp.279-286
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    • 1994
  • Fe(III) IDA -PEG/ dextran 액 이상 분계 시스템에서 phosphoprotein의 분배 예측을 위한 모델식이 유도되었다. 본 모델식에는 단백칠 분배에 수소이온과 hydroxide 이온의 저해효과가 포함되어 었다. 표변에 각각 1,27~의 phosphoryl 그 룹을 갖고 있는 ovalbumin의 AI, A2, 성분을 정제한 후 단백질 분배 실험을 수행하였으며 실 험결과와 모텔식에의한 예측이 잘 얼치하는 것 을 확인하였다. Fe(III)IDA-PEG와 단백질 표면 p phosphoryl group과의 결합상수는 PEG 상과 dextran상에 셔 각각 $6.1{\times}, 10^3M^{-1} and 2.3{\times}10^4M^{-1}$이었으며,이것은 Cu(II)IDA-PEG와 단백질 표면 histidine과의 결합상수 보다 3-5배 높은 값이 였 다.

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