• Title/Summary/Keyword: variety affinity

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An Approach to Isolation of Thromboxane Synthase (TX-SYN) by Ligand Tethered Affinity Techniques

  • Andersen Niels H.;Rhee Jaekeol
    • Bulletin of the Korean Chemical Society
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    • v.13 no.2
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    • pp.119-122
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    • 1992
  • The affinity chromatographic technique was applied to the isolation of Thromboxane Synthase, with a variety of imidazolyl alkanoic acids coupled Sepharose 2B including a gel (G in Table 4) which has one free COOH group in the bound affinity ligand. The effect of ligand structure on the "affinity" and "selectivity" for thromboxane synthase isolation is described.

Affinity and Variety between Words in the Framework of Hypernetwork (하이퍼네트워크에서 본 단어간 긴밀성과 다양성)

  • Kim, Joon-Shik;Park, Chan-Hoon;Lee, Eun-Seok;Zhang, Byoung-Tak
    • Journal of KIISE:Computer Systems and Theory
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    • v.35 no.4
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    • pp.166-171
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    • 2008
  • We studied the variety and affinity between the successive words in the text document A number of groups were defined by the frequency of a following word in the whole text (corpus). In the previous studies, the Zipf's power law was explained by Chinese restaurant process and hub node was searched after by examining the edge number profile in scale free network. We have observed both a power law and a hub profile at the same time by studying the conditional frequency and degeneracy of a group. A symmetry between the affinity and the variety between words were found during the data analysis. And this phenomenon can be explained within a viewpoint of "exploitation and exploration." We also remark on a small symmetry breaking phenomenon in TIPSTER data.

Functional Display of Maackia amurensis Hemagglutinin (MAH) on Bacteriophage (박테리오파아지 표면 발현 시스템을 이용한 Maackia amurensis Hemagglutinin (MAH)의 기능적 발현)

  • 임미정
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.176-179
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    • 2003
  • A library of unlimited number of novel lectins with diverse specificities has been previously generated by randomly mutating the carbohydrate-recognition domain of Maackia amurensis hemagglutinin (MAH). To establish the experimental environment capable of selecting high affinity mutant lectins in E. coli, phage display system was adapted. Carbohydrate binding capacity of two phagemid vectors, pComb3 and pComb8 displaying wild-type MAH lectin was assessed. Specific bindings of pComb3 and pComb8 phages expressing w.t. MAH to affinity-purified polyclonal anti-MAH antibody and to glycophorin was demonstrated. Both phages also showed strong hemagglutinating activity to intact but not sialidase-treated human erythrocytes, which is consistent to the specificity of native MAH. Taken together, two different phage display vectors successfully allowed the expression of active MAH as a fusion protein on the surface of bacteriophage, which will lead to preparation of unique plant lectins with high affinity toward a variety of carbohydrate chains.

Purification of Deoxycytidine Kinase from Various Human Leukemic Cells by End-product Analog Affinity Chromatography

  • Kim, Min-Young
    • BMB Reports
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    • v.28 no.4
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    • pp.281-289
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    • 1995
  • Homogeneous human deoxycytidine kinase was purified in one step from a variety of spontaneous human leukemic cells (T-ALL, B-ALL, B-CLL, AML, CML), and from cultured T-lymphoblast cells (MOLT-4) using the newly developed affinity medium, $dCp_4$-Sepharose. Starting with an ammonium sulfate fraction, purification was achieved in one step with the kinase being eluted from a column by the end product inhibitor, dCTP. The purified deoxycytidine kinase from T-ALL cells phosphorylated deoxyadenosine and deoxyguanosine, as well as deoxycytidine. The enzyme purified from T-ALL and B-CLL cells yielded one major band with a molecular weight of 52 kDa determined by SDS-polyacrylamide gel electrophoresis. AML and CML cells yielded one 52 kDa band and an extra band of 30 kDa molecular weight. On the other hand, B-ALL and MOLT-4 cells showed a low molecular weight band of 30 kDa only. However, the electrophoretic mobilities of enzymatic activity in 12% non-denaturing gels were identical for the dCyd kinase from all different kinds of leukemic cell lines, except that the B-ALL, B-CLL, and MOLT-4 cell preparations had an extra minor peak, all at the same position. dAdo and dCyd phosphorylating activities comigrated indicating that these activities are all associated with the same protein. Two new methods, a disk implantation method and a nitrocellulose powder method were used with a small amount of enzyme protein to raise polyclonal antibodies against dCyd kinase purified from T-ALL cells.

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

  • Chung, Bong-Hyun;Park, Young-Hoon
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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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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Structural and Thermodynamic Characteristics of cHLH Peptide and cHLH/HDM2 Complex

  • Im, Haeri;Cho, Sunhee;Ham, Sihyun
    • Proceeding of EDISON Challenge
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    • 2016.03a
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    • pp.62-66
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    • 2016
  • Tumor suppressor protein p53 loses its function upon binding with the HDM2 protein, and inhibiting the p53-HDM2 interaction is critical to suppress tumor cell growth. Recently, the cyclized helix-loop-helix peptide (cHLH) mimicking the ${\alpha}-helix$ part of the p53 protein has been designed and found to exhibit high binding affinity with HDM2. Here, we report the structural and thermodynamic characteristics of the bound complex of the cHLH peptide with the HDM2 protein. We performed molecular dynamics simulations to investigate the structural features of the cHLH peptide as well as its complex with the HDM2. The binding free energy calculation based on the integral equation theory was also executed to quantify the binding affinity for the cHLH/HDM2 complex and to understand the factors contributing to the binding affinity. We found a variety of factors for the helix stability of the cHLH peptide as well as in the complexation with the HDM2, which may provide an insight into the development of anti-cancer drug designs.

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Evaluation of proteomic strategies for analyzing ubiquitinated proteins

  • Peng, Jun Min
    • BMB Reports
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    • v.41 no.3
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    • pp.177-183
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    • 2008
  • Ubiquitin is an essential, highly-conserved small regulatory protein in eukaryotic cells. It covalently modifies a wide variety of targeted proteins in the forms of monomer and polymers, altering the conformation and binding properties of the proteins and thus regulating proteasomal delivery, protein activities and localization. Mass spectrometry has emerged as an indispensable tool for in-depth characterization of protein ubiquitination. Ubiquitinated proteins in cell lysates are usually enriched by affinity chromatography and subsequently analyzed by mass spectrometry for identification and quantification. Ubiquitin-conjugated amino acid residues can be determined by unique mass shift caused by the modification. Moreover, the complex structure of polyubiquitin chains on substrates can be dissected by bottom-up and middle-down mass spectrometric approaches, revealing potential novel functions of polyubiquitin linkages. Here I review the advances and caveats of these strategies, emphasizing caution in the validation of ubiquitinated proteins and in the interpretation of raw data.

Selective Concentration and Determination of Phenol and Chlorophenols by Solid Phase Extraction

  • Jung, Min-Woo;Lee, Dai Woon;Paeng, Ki-Jung
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.535-538
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    • 1995
  • This study employs the variety of mixtures of XAD resin and active carbons as concentration base for solid phase extraction (SPE) which has been widely used to preconcentrate and purify phenol and chlorophenols in determination of environmental water samples. In this study, we employed variety of mixtures of copolymer based XAD-4 resin with active carbons. This cartridges shows advantages of both materials, such as better affinity to phenol by active carbon and better mechanical stabilities from XAD resin. The better enrichment factor, pretreatment time, recoveries and limit of detection (LOD) were achieved by the attempts to pack precolumns with both meterials for preconcentration.

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An Extraction and Analysis of Problem Areas for BTL Projects from the Practical Perspective (실무적 관점에서의 BTL사업 문제영역 도출 및 분석)

  • Kim, Soo-Yong;Son, Myung-Chan;Yang, Jin-Kook
    • Korean Journal of Construction Engineering and Management
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    • v.14 no.3
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    • pp.157-166
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    • 2013
  • Unlike conventional BTO(Build-Transfer-Operate) projects, this BTL project aims at fixing the financial deficiencies of the government and expanding infrastructure through private capital. It allows the government to attract private capital for the construction of public facilities such as schools and social welfare agencies for whom private users don't need to pay, thereby bolstering national finance. BTL projects are causing a variety problems in progress. Therefore, we are required a practical approach to can improving a variety problems. In this study, we were derive the problem areas as follow. First, research data on the problem of domestic BTL projects, Second, high-performing foreign data analysis than domestic. And, we were analyzed to systematic management method for problem areas by using affinity techniques, matrix comparison analysis, AHP technique. Results of this study are expected provide management standards through real problem areas and the analyzing of relatively importance.