• Title/Summary/Keyword: Binding Ability

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One-Step Selection of Artificial Transcription Factors Using an In Vivo Screening System

  • Bae, Kwang-Hee;Kim, Jin-Soo
    • Molecules and Cells
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    • v.21 no.3
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    • pp.376-380
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    • 2006
  • Gene expression is regulated in large part at the level of transcription under the control of sequence-specific transcriptional regulatory proteins. Therefore, the ability to affect gene expression at will using sequencespecific artificial transcription factors would provide researchers with a powerful tool for biotechnology research and drug discovery. Previously, we isolated 56 novel sequence-specific DNA-binding domains from the human genome by in vivo selection. We hypothesized that these domains might be more useful for regulating gene expression in higher eukaryotic cells than those selected in vitro using phage display. However, an unpredictable factor, termed the "context effect", is associated with the construction of novel zinc finger transcription factors--- DNA-binding proteins that bind specifically to 9-base pair target sequences. In this study, we directly selected active artificial zinc finger proteins from a zinc finger protein library. Direct in vivo selection of constituents of a zinc finger protein library may be an efficient method for isolating multi-finger DNA binding proteins while avoiding the context effect.

The Potentiometric Performances of the Cation Selective Electrodes based on Tetracycline and Chemically Modified Tetracycline

  • Kang, Sang-Hyuk;Rhee, In-Sook;Paeng, Ki-Jung
    • Journal of Electrochemical Science and Technology
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    • v.2 no.3
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    • pp.143-145
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    • 2011
  • Metal-binding antibiotics are very attractive choices as cation selective ionophores. The ability of tetracycline (TC) antibiotics to bind to metal ions has obtained much attention. TCs exhibit the potentiometric performance changes for various cations dependant on several experiment conditions. In this report, we investigated the potentiometric performance changes of TC as the modification of TC's possible metal binding site. We found that the selectivity alter with the blocking main binding site of ionophores for cations. And, additionally it is possible to control the selectivity of sensors with chemical modification of ionophores.

Alkali Metal Cation Selectivity of [$1_7$]Ketonand in Methanol: Free Energy Perturbation and Molecular Dynamics Simulation Studies

  • 황선구;장윤희;유진하;정두수
    • Bulletin of the Korean Chemical Society
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    • v.20 no.10
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    • pp.1129-1135
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    • 1999
  • Free energy perturbation and molecular dynamics simulations were carried out to investigate the relative binding affinities of [17] ketonand (1) toward alkali metal cations in methanol. The binding affinities of 1 toward the alkali metal cations were calculated to be in the order Li+ > Na+ > K+ > Rb+ > Cs+, whereas our recent theoretically predicted and experimentally observed binding affinities for [18]starand (2) were in the order K+ > Rb+ > Cs+ > Na+ > Li+. The extremely different affinities of 1 and 2 toward smaller cations, Li + and Na+ , were explained in terms of the differences in their ability to change the conformation to accommodate cations of different sizes. The carbonyl groups constituting the central cavity of 1 can reorganize to form a cavity with the optimal M+ -O distance, even for the smallest Li+, without imposing serious strain on 1. The highest affinity of 1 for Li+ was predominantly due to the highest Coulombic attraction between the smallest Li+ and the carbonyl oxygens of 1.

In Vitro Assays of Sperm Fertility (정자의 수정능력 평가기법)

  • 박수봉
    • Journal of Embryo Transfer
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    • v.10 no.1
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    • pp.15-22
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    • 1995
  • Since the turn of the century, scientists have earnestly sought to develop a single laboratory assay or combination of laboratory assays which accurately predict the fertility of a semen sample. Most of these assays have focused on evaluating physical characteristics of sperm such as motility, viability, acrosomal integrity and morphology. In recent years new approches have been used to assess the functional aspects of a sperm that are needed to reach the oocyte, fertilize it and contribute to successful embryo development. Among these techniques are the ability of sperm to undergo a heparin induced acrosome reaction and in vitro fertilization, and the affinity of sperm to bind heparin binding protein. Intensification of research efforts in the area of control of sperm fertilizing ability should be a high priority, in view of undoubted benifits both to our basic understanding of sperm fertilizing ability and to our ability to modify it for Al industry.

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Binding of $^3$H-Lectin from Kintoki Bean and Taro Tuber to Small Intestine of the Mouse (콩과 토란에서 추출한 $^3$H-Lectin의 마우스 소장에의 흡착량 정량)

  • 서영주
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.4
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    • pp.489-493
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    • 1993
  • The major objective of this study carried out was to compare the binding of Kintoki bean lectin (KBL) and Taro tuber lectin (TTL) to the mouse intestinal segments using $^3$H-labeled lectins and to assess the effect of such binding on the ability of the small intestine. Binding of $^3$H-KBL or $^3$H-TTL was studied under various conditions of time course, temperature, concentration, pH and additives of sugars, EDTA or unlabeled native lectin. The interaction of the lectins to intestinal tissue was stronger in KBL than in TTL, which was supposed to be the major reason for the stronger antinuritional enen of KBL. The optimal binding conditions were at 37$^{\circ}C$ for 60mins and at pH 7. The binding of both lectins were inhibited by fetuin and EDTA.

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Multiple hTAFII31-binding motifs in the intrinsically unfolded transcriptional activation domain of VP16

  • Kim, Do-Hyoung;Lee, Si-Hyung;Nam, Ki-Hoon;Chi, Seung-Wook;Chang, Ik-Soo;Han, Kyou-Hoon
    • BMB Reports
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    • v.42 no.7
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    • pp.411-417
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    • 2009
  • Transcriptional activation domain (TAD) in virion protein 16 (VP16) of herpes simplex virus does not have any globular structure, yet exhibits a potent transcriptional activity. In order to probe the structural basis for the transcriptional activity of VP16 TAD, we have used NMR spectroscopy to investigate its detailed structural features. Results show that an unbound VP16 TAD is not merely "unstructured" but contains four short motifs (residues 424-433, 442-446, 465-467 and 472-479) with transient structural order. Pre-structured motifs in other intrinsically unfolded proteins (IUPs) were shown to be critically involved in target protein binding. The 472-479 motif was previously shown to bind to $hTAF_{II}31$, whereas the $hTAF_{II}31$-binding ability of other motifs found in this study has not been addressed. The VP16 TAD represents another IUP whose pre-structured motifs mediate promiscuous binding to various target proteins.

Comparative Analysis on the Cytotoxicity of Naegleria fowleri and N. gruberi to Macrophages by the Addition of Saccharides

  • Jung, Suk-Yul
    • Biomedical Science Letters
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    • v.16 no.4
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    • pp.221-227
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    • 2010
  • To elucidate the invasion mechanism of pathogenic Naegleria fowleri, especially a receptor-ligand recognition, we investigated the in vitro cytotoxicity of pathogenic N. fowleri and nonpathogenic N. gruberi to murine macrophages, RAW 264.7, by adding four kinds of saccharides, ${\alpha}$-fucose, ${\beta}$-galactose, ${\alpha}$-D-mannopyranoside (${\alpha}$-mannose) and xylose. There was not enough of a difference in the cytotoxicity of N. fowleri treated with 10 mM of each saccharide. In particular, the cytotoxicity of N. fowleri was highly inhibited by 100 mM ${\alpha}$-mannose, which was 62.3% inhibition calculated by the analysis of lactate dehydrogenase (LDH) release assay. Although murine macrophages were not significantly destroyed by nonpathogenic N. gruberi under hematoxylin staining, the cytotoxicity of N. gruberi was inhibited from 31.5% to 14.5% (P<0.01) by 100 mM ${\alpha}$-mannose treatment. The binding of N. fowleri to macrophages was inhibited from 33% to 50% by 100 mM ${\alpha}$-mannose. Furthermore, as results of the adhesion assays which were performed to determine whether binding of Naegleria is mediated by saccharides-binding protein, the binding ability of N. fowleri as well as N. gruberi was inhibited by 100 mM ${\alpha}$-mannose.

Ginsenoside-Rb1 Acts as a Weak Estrogen Receptor Agonist Independent of Ligand Binding.

  • Park, Wan-Kyu;Jungyoon Cho;Lee, Young-Joo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.114-114
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    • 2003
  • Ginseng is a medicinal herb widely used in Asian countries, and its pharmacological effects has been demonstrated in various systems such as cardiovascular, central nervous, and endocrine systems. Its effects are mainly attributed to the ginsenosides. We hypothesize that a component of Panax ginseng, ginsenoside-Rbl, acts by binding to estrogen receptor. We have investigated the estrogenic activity of ginsenoside-Rbl in a transient transfection system using estrogen receptors ${\alpha}$ or ${\beta}$ with estrogen -responsive luciferase plasmids in COS monkey kidney cells. Ginsenoside-Rbl activated both estrogen receptors ${\alpha}$ and ${\beta}$ in a dose-dependent manner (0.5 -100 M ). Activation was inhibited by the specific estrogen receptor antagonist ICI 182,780, indicating that the estrogenic effect of ginsenoside-Rbl is estrogen receptor dependent. Next, we evaluated the ability of ginsenoside-Rbl to induce estrogen-responsive progesterone receptor gene by semi-quantitative RT-PCR assays. MCF-7 cells treated with l7${\beta}$-estradiol or ginsenoside- Rb1 exhibited an increased expression of progesterone receptor mRNA. However, ginsenoside-Rbl failed to displace the specific binding of [3H]17${\beta}$-estradiol to estrogen receptor in MCF-7 cells as examined by whole cell ligand binding assays, suggesting that there is no direct interaction of ginsenoside-Rbl with estrogen receptor. Our results indicate that estrogen-like activity of ginsenoside-Rbl is independent of direct estrogen receptor association.

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Studies on pathogenicity of Escherichia coli isolated from chickens: Correlation between in vitro Congo-red binding properties and in vivo virulence in avian Escherichia coli (닭유래 Escherichia coli의 병원성에 관한 연구 : 시험관내 Congo-red 결합능과 병원성간의 상관관계)

  • Woo, Yong Ku;Kim, Ki Seuk;Kim, Bong Hwan
    • Korean Journal of Veterinary Research
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    • v.31 no.1
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    • pp.55-61
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    • 1991
  • The correlation between in vitro Congo-red binding properties of E coli and in vivo invasiveness of the organisms in SPF chickens and mice was investigated. Congo-red positive E coli colonies were dark-red color with a typical colonial morphology of rough appearance when grown on Congo-red medium, while Congo-red negative colonies showed pale-pink color and smooth surfaced colonial morphology. Pathogenicity of 10 Congo-red positive E coli for mice was observed in 92.5% but that of 5 Congo-red negative E coli was 45%. Invasiveness of 10 Congo-red positive E coli for chickens was observed in 96% of the SPF chickens tested but that of 5 Congo-red negative E coli was 16% only. These results of pathogenicity studies with E coli isolates indicate a significant correlation between Congo-red binding ability and virulence in avian Escherichia coli.

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DNA-Binding and Thermodynamic Parameters, Structure and Cytotoxicity of Newly Designed Platinum(II) and Palladium(II) Anti-Tumor Complexes

  • Mansouri-Torshizi, Hassan;Saeidifar, Maryam;Khosravi, Fatemeh;Divsalar, Adeleh;Saboury, Ali.Akbar;Ghasemi, Zahra Yekke
    • Bulletin of the Korean Chemical Society
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    • v.32 no.3
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    • pp.947-955
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    • 2011
  • The complexes [Pd(bpy)(Hex-dtc)]$NO_3$ and [Pt(bpy)(Hex-dtc)]$NO_3$ (bpy is 2,2'-bipyridine and Hex-dtc is hexyldithiocarbamato ligands) were synthesized and characterized by elemental analysis and spectroscopic studies. The cytotoxicity assay of the complexes has been performed on chronic myelogenous leukemia cell line, K562, at micromolar concentration. Both complexes showed cytotoxic activity far better than that of cisplatin under the same experimental conditions. The binding parameters of the complexes with calf thymus DNA (CT-DNA) was investigated using UV-visible and fluorescence techniques. They show the ability of cooperatively intercalating in CT-DNA. Gel filtration studies demonstrated that platinum complex could cleave the DNA. In the interaction studies between the Pd(II) and Pt(II) complexes with CT-DNA, several binding and thermodynamic parameters have been determined, which may provide deeper insights into the mechanism of action of these types of complexes with nucleic acids.