• Title/Summary/Keyword: Binding Ability

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Effects of Thyroid Hormone on Pteroylpolyglutamate Chain Length and the Binding Activity of Folate Binding Protein in Rat Liver (갑상선 호르몬이 흰쥐 간세포내 엽산의 Polyglutamate 직쇄분포와 세포질 엽산 결합단백질의 결합성에 미치는 영향)

  • 민혜선
    • Journal of Nutrition and Health
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    • v.32 no.4
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    • pp.369-375
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    • 1999
  • Hyperthroidism in known to alter the activity of a number of enzymes involved in the catabolism of histidine to CO2. 10-Formyltetrahydrofolate dehydrogenase(EC 1.5, 1.6, 10-formyl-THE dehydrogenase) catalyzes the NADP-dependent conversion of 10-formyltetrahydrofolate to tetrahydrofolate and CO2. In previous studies, 10-formyl-THF dehydrogenase purified from rat and pig liver was coidentified with the cytosolic folate-binding protein. In this study, we investigated the effects of feeding thyroid powder (TP) and thiouracil(TU) on the folate-binding properties of 10-formyl-THE dehydrogenase, the uptake of an injected dose of [3H] folate, and the metabolism of labeled folate to pteroylopoly-${\gamma}$-glutamate in rat liver. The initial hepatic uptake(24hr) of the labeled folate dose was higher in TU-rats and slightly higher in TP-rats in controls. With longer time periods, decreased hepatic uptake of labeled folate was observed in TP-animals compared to euthroid animals, and high levels of hepatic uptake of labeled folate were maintained in TU-animals. This data shows that high levels of thyroid hormone decreased the retention of folate in rat liver. Folate polygutamate chain length was shorter in TU-rats than controls, which suggests that thyroid states do not affect the ability to synthesize pteroylpolyglutamates and that folate polyglutamate might be modulated by altered folate pool size. The ability of 10-formyl-THE dehydrogenase to bind folate in rat liver was similar in both TP-and TU-rats although dehydrogenase activity was changed by thyroid sates.

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Label-free Detection of the Transcription Initiation Factor Assembly and Specific Inhibition by Aptamers

  • Ren, Shuo;Jiang, Yuanyuan;Yoon, Hye Rim;Hong, Sun Woo;Shin, Donghyuk;Lee, Sangho;Lee, Dong-Ki;Jin, Moonsoo M.;Min, Irene M.;Kim, Soyoun
    • Bulletin of the Korean Chemical Society
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    • v.35 no.5
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    • pp.1279-1284
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    • 2014
  • The binding of TATA-binding protein (TBP) to the TATA-box containing promoter region is aided by many other transcriptional factors including TFIIA and TFIIB. The mechanistic insight into the assembly of RNA polymerase II preinitation complex (PIC) has been gained by either directly altering a function of target protein or perturbing molecular interactions using drugs, RNAi, or aptamers. Aptamers have been found particularly useful for studying a role of a subset of PIC on transcription for their ability to inhibit specific molecular interactions. One major hurdle to the wide use of aptamers as specific inhibitors arises from the difficulty with traditional assays to validate and determine specificity, affinity, and binding epitopes for aptamers against targets. Here, using a technique called the bio-layer interferometry (BLI) designed for a label-free, real-time, and multiplexed detection of molecular interactions, we studied the assembly of a subset of PIC, TBP binding to TATA DNA, and two distinct classes of aptamers against TPB in regard to their ability to inhibit TBP binding to TFIIA or TATA DNA. Using BLI, we measured not only equilibrium binding constants ($K_D$), which were overall in close agreement with those obtained by electrophoretic mobility shift assay, but also kinetic constants of binding ($k_{on}$ and $k_{off}$), differentiating aptamers of comparable KDs by their difference in binding kinetics. The assay developed in this study can readily be adopted for high throughput validation of candidate aptamers for specificity, affinity, and epitopes, providing both equilibrium and kinetic information for aptamer interaction with targets.

Heterocyclic Amines Removal by Binding Ability of Lactic Acid Bacteria Isolated from Soybean Paste (된장에서 분리된 유산균의 결합력에 의한 Heterocyclic Amines 제거)

  • Lim, Sung-Mee
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.73-83
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    • 2014
  • The objective of the this study was to investigate the binding capacity and removal ability of lactic acid bacterial strains obtained from Korean soybean paste for mutagenic heterocyclic amines (HCAs) formed during cooking of protein-rich food at high temperature. Among 19 strains identified by carbohydrate fermentation and 16S rRNA sequencing, the live cell or cell-free culture supernatant of Lactobacillus acidophilus D11, Enterococcus faecium D12, Pediococcus acidilactici D19, L. acidophilus D38, Lactobacillus sakei D44, Enterococcus faecalis D66, and Lactobacillus plantarum D70 inhibited the mutagenesis caused by either 3-amino-1,4-dimethyl-5H-pyrido[4,3-b] indole (Trp-P-1) or 3-amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) in Salmonella typhimurium TA98 and TA100. The bacterial cells of the isolated strains showed greater binding activity than the pure cell wall, exopolysaccharide, and pepetidoglycan. The carbohydrate moieties of the cell wall or protein molecules on the cell surface have a significant role in binding Trp-P-1 and Trp-P-2, since protease, heating, sodium metaperiodate, or acidic pH treatments significantly (P<0.05) reduced the binding efficacy of the tested bacteria. Addition of metal ions or sodium dodecyl sulfate decreased the binding ability of E. faecium D12, L. acidophilus D38, and E. faecalis D66. Therefore, the binding mechanisms of these strains may consist of ion-exchange and hydrophobic bonds. Especially, the high mutagen binding by L. acidophilus D38 and L. plantarum D70 may reduce the accumulation or absorption of Trp-P-1 and Trp-P-2 in the small intestine via increased excretion of a mutagen-bacteria complex.

The Roles of Tryptophan and Histidine Residues in the Catalytic Activities $\beta$-Cyclodextrin Glucanotransferase from Bacillus firmus var. alkalophilus

  • Shin, Hyun-Dong;Kim, Chan;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.62-69
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    • 1999
  • In order to investigate the critical amino acid residues involved in the catalytic activities of $\beta$-cyclodextrin glucanotransferase ($\beta$-CGTase) excreted by Bacillus firmus var. alkalophilus, the amino acid residues in $\beta$-CGTase were modified by various site-specific amino acid modifying reagents. The cyclizing and amylolytic activities of $\beta$-CGTase were all seriously reduced after treatment with Woodward's reagent K (WRK) modifying aspartic/glutamic acid, N-bromosuccinimde (NBS) modifying tryptophan, and diethylpyrocarbonate (DEPC) modifying histidine residues. The roles of tryptophan and histidine residues in $\beta$-CGTase were further investigated by measuring the protection effect of various substrates during chemical modification, comparing protein mobility in native and affinity polyacrylamide gel electrophoresis containing soluble starch, and comparing the $K_m$ and $V_{max}$ values of native and modified enzymes. Tryptophan residues were identified as affecting substrate-binding ability rather than influencing catalytic activities. On the other hand, histidine residues influenced catalytic ability rather than substrate-binding ability, plus histidine modification had an effect on shifting the optimum pH and pH stability.

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A Comparative Study of the Influence of Miflumic Acid and Phenylbutazone on Warfarin-Plasma Protein Binding (약-약 상호작용 연구(IV) Warfarin의 혈장단백 결합에 대한 Niflumic Acid 및 Phenylbutazone의 영향 비교)

  • 조윤성;양중익
    • YAKHAK HOEJI
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    • v.24 no.2
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    • pp.97-100
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    • 1980
  • To determine in vitro effects of phenylbutazone and niflumic acid on warfarin binding to rabbit serum protein, warfarin was added to the rabbit plasma, and the bound fraction was determined by warfarin-protein complex fluorescence. The bound fraction was decreased by phenylbtazone and niflumic acid. From this effect niflumic acid was found to have the more potent ability to displace warfarin from protein binding sites than phenylbutazone.

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STRUCTURAL ANALYSIS OF RAPAMYCIN'S ROLE IN BINDING FKBP12 AND FRAP

  • Park, Jungwon;Jie Chen;Stuart Schreiber;Jon Clardy
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.9-9
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    • 1996
  • The immunosuppressive and cell cycle arrest agent rapamycin works by binding together two proteins: the FK506 binding protein (FKBP12) and the FKBP-rapamycin associated protein (FRAP). A 2.7 $\AA$ resolution crystal structure of the triple complex of human FK506 binding protein (FKBP12), rapamycin, and FKBP12-rapamycin binding domain (FRB) of FRAP, reveals two proteins bound together through rapamycin' s ability to simultaneously occupy two different hydrophobic binding pockets. (omitted)

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Changes in the Allergenicity of Saeujeot by Fermentation

  • Kim, Seong-Mi;Park, Jin-Gyu;Kim, Koth-Bong-Woo-Ri;Saeki, Hiroki;Nakamura, Atsushi;Lee, Ju-Woon;Byun, Myung-Woo;Ahn, Dong-Hyun
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.919-924
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    • 2008
  • The aim of this study was to observe the changes in allergenicity of saeujeot (salted and fermented shrimp) using a competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). The fermentation conditions tested for saeujeot consisted of various temperatures (25, 15, and $5^{\circ}C$) and salt concentrations (25, 15, and 10%). When saeujeot was fermented at a low salt concentration and high temperature, the binding ability of mAb and shrimp-allergic patient serum to allergen was significantly decreased. In particular, the binding ability of mAb to allergen in saeujeot fermented with 10% salt at $25^{\circ}C$ for 5 days decreased to 5%. Also, the binding ability of shrimp-allergic patient serum to allergen in saeujeot fermented for 5 days with 10% salt at $25^{\circ}C$ was 8%. In conclusion, the binding of mAb and shrimp-allergic patient serum to tropomyosin in saeujeot decreased with longer fermentation periods, lower salt concentrations (10%), and higher temperatures ($25^{\circ}C$).

The Synthetic Melanin Nanoparticles Having An Excellent Binding Capacity of Heavy Metal Ions

  • Kim, Da Jeong;Ju, Kuk-Youn;Lee, Jin-Kyu
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3788-3792
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    • 2012
  • Spherical-shape melanin nanoparticles with good water-dispersibility were successfully synthesized by a simple oxidation polymerization of 3,4-dihydroxy-phenylalanin (DOPA) with $KMnO_4$. Similar features to those known from natural and synthetic melanin polymers were observed from prepared melanin nanoparticles by FT-IR, UV-Vis., and ESR spectroscopic methods. Their binding ability with several heavy metal ions from aqueous solution was quantitatively investigated, and the maximum binding capacities with melanin nanoparticles to lead, copper, and cadmium ions were obtained as 2.45, 2.17 and 1.88 mmol/g, respectively, which are much larger values than those reported from natural and synthetic melanin polymers. The large binding capacity and fast binding rate of melanin nanoparticles to metal ions can make them an excellent candidate for the remediation of contaminated water.

Engineering and Application of Zinc Finger Proteins and TALEs for Biomedical Research

  • Kim, Moon-Soo;Kini, Anu Ganesh
    • Molecules and Cells
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    • v.40 no.8
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    • pp.533-541
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    • 2017
  • Engineered DNA-binding domains provide a powerful technology for numerous biomedical studies due to their ability to recognize specific DNA sequences. Zinc fingers (ZF) are one of the most common DNA-binding domains and have been extensively studied for a variety of applications, such as gene regulation, genome engineering and diagnostics. Another novel DNA-binding domain known as a transcriptional activator-like effector (TALE) has been more recently discovered, which has a previously undescribed DNA-binding mode. Due to their modular architecture and flexibility, TALEs have been rapidly developed into artificial gene targeting reagents. Here, we describe the methods used to design these DNA-binding proteins and their key applications in biomedical research.