• Title/Summary/Keyword: Binding Ability

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Characterization of Binding of Treponema denticola to Immobilized Fibrinogen using the Fluorescent Fatty Acid Labeling Method

  • Hong, Jin;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.35 no.3
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    • pp.107-111
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    • 2010
  • Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

Transcriptional Response and Enhanced Intestinal Adhesion Ability of Lactobacillus rhamnosus GG after Acid Stress

  • Bang, Miseon;Yong, Cheng-Chung;Ko, Hyeok-Jin;Choi, In-Geol;Oh, Sejong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.10
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    • pp.1604-1613
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    • 2018
  • Lactobacillus rhamnosus GG (LGG) is a probiotic commonly used in fermented dairy products. In this study, RNA-sequencing was performed to unravel the effects of acid stress on LGG. The transcriptomic data revealed that the exposure of LGG to acid at pH 4.5 (resembling the final pH of fermented dairy products) for 1 h or 24 h provoked a stringent-type transcriptomic response wherein stress response- and glycolysis-related genes were upregulated, whereas genes involved in gluconeogenesis, amino acid metabolism, and nucleotide metabolism were suppressed. Notably, the pilus-specific adhesion genes, spaC, and spaF were significantly upregulated upon exposure to acid-stress. The transcriptomic results were further confirmed via quantitative polymerase chain reaction analysis. Moreover, acid-stressed LGG demonstrated an enhanced mucin-binding ability in vitro, with 1 log more LGG cells (p < 0.05) bound to a mucin layer in a 96-well culture plate as compared to the control. The enhanced intestinal binding ability of acid-stressed LGG was confirmed in an animal study, wherein significantly more viable LGG cells (${\geq}2log\;CFU/g$) were observed in the ileum, caecum, and colon of acid-stressed LGG-treated mice as compared with a non-acid-stressed LGG-treated control group. To our knowledge, this is the first report showing that acid stress enhanced the intestine-binding ability of LGG through the induction of pili-related genes.

Functional Characteristics of C-terminal Lysine to Cysteine Mutant Form of CTLA-4Ig

  • Kim, Bongi;Shin, Jun-Seop;Park, Chung-Gyu
    • IMMUNE NETWORK
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    • v.13 no.1
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    • pp.16-24
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    • 2013
  • CTLA-4Ig is regarded as an inhibitory agent of the T cell proliferation via blocking the costimulatory signal which is essential for full T cell activation. To improve applicability, we developed the CTLA-4Ig-CTKC in which the c-terminal lysine had been replaced by cysteine through single amino acid change. The single amino acid mutation of c-terminus of CTLA-4Ig was performed by PCR and was checked by in vitro transcription and translation. DNA construct of mutant form was transfected to Chinese hamster ovary (CHO) cells by electroporation. The purified proteins were confirmed by Western blot and B7-1 binding assay for their binding ability. The suppressive capacity of CTLA-4Ig-CTKC was evaluated by the mixed lymphocyte reaction (MLR) and in the allogeneic pancreatic islet transplantation model. CTLA-4Ig-CTKC maintained binding ability to B7-1 molecule and effectively inhibits T cell proliferation in MLR. In the murine allogeneic pancreatic islet transplantation, short-term treatment of CTLA-4Ig-CTKC prolonged the graft survival over 100 days. CTLA-4Ig-CTKC effectively inhibits immune response both in MLR and in allogeneic islet transplantation model, indicating that single amino acid mutation does not affect the inhibitory function of CTLA-4Ig. CTLA-4Ig-CTKC can be used in vehicle-mediated drug delivery system such as liposome conjugation.

Water Binding Capacity of Vegetable Fiber (식이섬유의 수분결합력에 관한 연구)

  • 계수경
    • The Korean Journal of Food And Nutrition
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    • v.9 no.3
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    • pp.231-235
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    • 1996
  • The water binding capacity(WC) of acid detergent fiber(ADF) was estimated. The WBC of raw vegetables ranged from 5. 1g to 24.7g water/g ADF. WBC was high in pepers and low in welsh onions. The correlation coefficients between WBC and fiber components were examined to fled which component is responsible for the deteruuning ability to bind water. The correlation coefficient between WBC and cellulose was +0.8. The binding capacity of water by fiber was affected positively by cellulose. fermentation increased in WBC of ADF. Changes of no in accordance with pH changes were evaluated at pH 2, 5.2 and 6. In all cases, WBC was high In weak acid and neutral.

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Enhancement of Calcium-Binding Quality of Proglycinin Peptides by Chemical Phosphorylation

  • Yang, Jung-Ik;Lee, Shin-Hee;Hahm, Dae-Hyun;Kim, Il-Hwan;Choi, Sang-Yun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.607-611
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    • 2004
  • Glycinin, one of the predominant storage proteins in soybeans, was examined as to whether it could be used as a calcium-binding mediator after chemical phosphorylation and enzymatic hydrolysis. Glycinin is composed of six subunits. One of the proglycinin subunits $(A_{la}B_{lb})$ was overexpressed in E. coli to obtain nonphosphorylated proteins with homogeneity. To investigate the enhanced calcium-binding properties of the phosphopeptides, the proglycinin was purified, phosphorylated, and hydrolyzed with trypsin. The proglycinin expressed in E. coli was purified by ammonium sulfate precipitation, ion-exchange chromatography, and cryoprecipitation. Chemical phosphorylation by sodium trimetaphosphate was performed to obtain phosphorylated proglycinin. After the phosphorylation, one-dimensional isoelectric focusing gel electroanalysis confirmed the phosphorylation of the proglycinin. The phosphorylated peptides were then hydrolyzed with trypsin, followed by a binding reaction with calcium chloride. The calcium-bound phosphopeptides were finally separated using immobilized metal $(Ca^{2+})$ chromatography. Consequently, a limited tryptic hydrolysate of the isolated phosphopeptides exhibited an enhanced calcium-binding ability, suggesting the potential of glycinin phosphopeptides as a calcium-binding mediator with greater availability.

Antibacterial Activity of Essential Oils on the Growth of Staphylococcus aureus and Measurement of their Binding Interaction Using Optical Biosensor

  • Chung, Kyong-Hwan;Yang, Ki-Sook;Kim, Jin;Kim, Jin-Chul;Lee, Ki-Young
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1848-1855
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    • 2007
  • Antibacterial activity of essential oils (Tea tree, Chamomile, Eucalyptus) on Staphylococcus aureus growth was evaluated as well as the essential oil-loaded alginate beads. The binding interactions between the cell and the essential oils were measured using an optical biosensor. The antibacterial activity of the essential oils to the cell was evaluated with their binding interaction and affinity. The antibacterial activity appeared in the order of Tea Tree>Chamomile>Eucalyptus, in comparison of the inhibition effects of the cell growth to the essential oils. The association rate constant and affinity of the cell binding on Tea Tree essential oil were $5.0{\times}10^{-13}\;ml/(CFU{\cdot}s)$ and $5.0{\times}10^5\;ml/CFU$, respectively. The affinity of the cell binding on Tea Tree was about twice higher than those on the other essential oils. It might be possible that an effective antibacterial activity of Tea Tree essential oil was derived from its strong adhesive ability to the cell, more so than those of the other essential oils.

In Vitro Estrogenic Activity of Silkworm (Bombyx mon) Pupa and Herbs (누에(Bombyx mori) 번데기 및 한약재의 In Vitro 에스트로젠 활성)

  • Yang Ji-Won;Choi Eun-Mi;Kwon Mu-Gil;Koo Sung-Ja
    • Journal of the East Asian Society of Dietary Life
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    • v.15 no.3
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    • pp.315-322
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    • 2005
  • In this study we report on the estrogen activity of silkworm pupa and herb extracts in vitro. The estrogenic activity of these resources was investigated by competition binding assays with estrogen receptor $\alpha(ER{\alpha})\;or\;ER{\beta}$, and viability of MCF-7 cells, a human breast cancer cell line. Saturation ligand-binding analysis of $ER{\alpha}\;and\;ER{\beta}$ revealed that all plant extracts competed with estrogen ligand for binding to both ER subtypes with a similar preference and degree and competed stronger with ligand for binding to $ER{\beta}\;than\;to\;ER{\alpha}$. The highest $ER{\alpha}-binding$ sample was silkworm pupa aqueous extract The highest $ER{\beta}-binding$ sample was silkworm pupa oil. These samples were further tested for bioactivity based on their ability to regulate cell growth rate in ER(+) breast cancer cell line, MCF-7 cells. Our studies showed that silkworm pupa, soritae, sesame, yam, pueraria, malt, ginseng, Polygonum multiflorum, and Curcuma longa significantly stimulated the growth of MCF-7 cells (P<0.05). In summary, these results suggested that silkworm pupa and herbs might be useful as potential phytoestrogens.

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RNA Binding Protein-Mediated Post-Transcriptional Gene Regulation in Medulloblastoma

  • Bish, Rebecca;Vogel, Christine
    • Molecules and Cells
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    • v.37 no.5
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    • pp.357-364
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    • 2014
  • Medulloblastoma, the most common malignant brain tumor in children, is a disease whose mechanisms are now beginning to be uncovered by high-throughput studies of somatic mutations, mRNA expression patterns, and epigenetic profiles of patient tumors. One emerging theme from studies that sequenced the tumor genomes of large cohorts of medulloblastoma patients is frequent mutation of RNA binding proteins. Proteins which bind multiple RNA targets can act as master regulators of gene expression at the post-transcriptional level to co-ordinate cellular processes and alter the phenotype of the cell. Identification of the target genes of RNA binding proteins may highlight essential pathways of medulloblastomagenesis that cannot be detected by study of transcriptomics alone. Furthermore, a subset of RNA binding proteins are attractive drug targets. For example, compounds that are under development as anti-viral targets due to their ability to inhibit RNA helicases could also be tested in novel approaches to medulloblastoma therapy by targeting key RNA binding proteins. In this review, we discuss a number of RNA binding proteins, including Musashi1 (MSI1), DEAD (Asp-Glu-Ala-Asp) box helicase 3 X-linked (DDX3X), DDX31, and cell division cycle and apoptosis regulator 1 (CCAR1), which play potentially critical roles in the growth and/or maintenance of medulloblastoma.

가상 소화모델시스템 내에서 감마선 조사와 가열 처리된 Ovalbumin의 IgE-Binding Ability의 변화

  • Seo, Ji-Hyeon;Kim, Jae-Hun;O, Sang-Hui;Lee, Ju-Un;Byeon, Myeong-U
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.10a
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    • pp.412-415
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    • 2004
  • 본 연구는 감마선 조사를 이용한 알레르기 저감화 식품을 개발하기 위한 실용화 방법의 안전성을 검토하기 위해 실시되었다. 감마선 조사 후 가열 처리된 OVA는 전기영동상에서pepsin에 의해서 상당히 감소하는 것을 확인할 수 있었다. IgG 항체를 통해 살펴본 소화성은 intact OVA보다 증가하여 가수분해되기 쉬운 형태임을 알 수 있었다. 또한 효소처리 전의 IgE-binding ability는 현저히 감소하였고 최종 효소분해까지 일정하게 낮은 농도가 유지되어 가상 소화모델 시스템 내에서의 안전성을 확인할 수 있었다. 이상의 결과에서 감마선 및 가열 처리를 단독으로 시행하는 공정보다는 감마선과 가열을 병용하는 방법이 알레르기 저감 식품을 개발에 더욱 효과적일 것으로 사료된다.

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