• Title/Summary/Keyword: Binding study

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Surface interactions between two of the main periodontal pathogens: Porphyromonas gingivalis and Tannerella forsythia

  • Zhu, Weidong;Lee, Seok-Woo
    • Journal of Periodontal and Implant Science
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    • v.46 no.1
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    • pp.2-9
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    • 2016
  • Purpose: Porphyromonas gingivalis and Tannerella forsythia have been implicated as the major etiologic agents of periodontal disease. These two bacteria are frequently isolated together from the periodontal lesion, and it has been suggested that their interaction may increase each one's virulence potential. The purpose of this study was to identify proteins on the surface of these organisms that are involved in interbacterial binding. Methods: Biotin labeling of surface proteins of P. gingivalis and T. forsythia and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was performed to identify surface proteins involved in the coaggregating activity between P. gingivalis and T. forsythia. Results: It was found that three major T. forsythia proteins sized 161, 100, and 62 kDa were involved in binding to P. gingivalis, and P. gingivalis proteins sized 35, 32, and 26 kDa were involved in binding to T. forsythia cells. Conclusions: LC-MS/MS analysis identified one T. forsythia surface protein (TonB-linked outer membrane protein) involved in interbacterial binding to P. gingivalis. However, the nature of other T. forsythia and P. gingivalis surface proteins identified by biotin labeling could not be determined. Further analysis of these proteins will help elucidate the molecular mechanisms that mediate coaggregation between P. gingivalis and T. forsythia.

Differential Expression of Cyclic AMP-Response Element Binding Protein Zhangfei (CREBZF) in the Mouse Testis during Postnatal Development

  • Jang, Hoon
    • Development and Reproduction
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    • v.22 no.1
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    • pp.65-72
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    • 2018
  • Cyclic AMP-response element binding protein zhangfei (CREBZF), a member of ATF/CREB (activating transcription factor/ cAMP response element binding protein) family, regulates numerous cellular functions and development of cells by interacting transcription factors. This study discovered the expression pattern of CREBZF in seminiferous tubule of testes during the postnatal development of mice. In testis, CREBZF mRNA expression was the highest among other organs. Immunofluorescence analyses showed that the CREBZF was specifically expressed on spermatocyte but not in spermatogonia and Sertoli cells in seminiferous epithelium of mouse testis. Semi-quantitative polymerase chain reaction (PCR) analysis showed that CREBZF transcript level was significantly elevated during postnatal development of mouse testis. Confocal imaging analysis indicated that the protein expression of CREBZF in seminiferous tubule remained low until postnatal day (PD) 14, and was dramatically increased in PD 21. Interestingly, only one type of the spermatocyte expressed CREBZF specifically among SCP3-positive spermatocytes. Taken together, these results suggest that CREBZF may be novel putative marker of the spermatocyte and regulate meiosis during postnatal development of mice.

NMR Studies of Zinc-binding Luteinizing Hormone Releasing Hormone

  • Kim, Dae-Sung;Lee, Mi-Sun;Lee, Chang-Jun;Won, Ho-Shik
    • Journal of the Korean Magnetic Resonance Society
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    • v.10 no.2
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    • pp.163-174
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    • 2006
  • Luteinizing Hormone Releasing Hormone(LHRH) is a decapeptide neurotransmitter known to be regulated by metal ions in the hyperthalamus. Zn-binding LHRH complex was systhesized, and zinc-LHRH complex was studied to understand what kinds of structural modifications would be critical in the LHRH releasing mechanism. Both nonexchangeable and exchangeable $^1H-NMR$ signal assignments were accomplished by pH-dependent and COSY NMR experiments. In addition, $^1H-NMR$ chemical shift changes of a-proton and peptide NH NMR signals at different pH condition, and $^1H-NMR$ signal differences between metal free and metallo-LHRH complex was monitored. NMR signals exhibit that primary metal-binding sites are nitrogens donor of imidazole ring and Arg, and peptide oxygen of Pro-His in the sequence. Structure obtained in this study has a cyclic conformation which is similar to that of energy minimized, and exhibits a specific a-helical turn with residue numbers $(2{\sim}7)$ out of 10 amino acids.

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Changes of the Binding Abilities of Immunoglobulin G and E on Gamma-Irradiated Ovalbumin by Proteolytic Enzymes

  • Lee, Ju-Woon;Seo, Ji-Hyun;Kim, Jae-Hun;Yook, Hong-Sun;Lee, Soo-Young;Byun, Myung-Woo
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.355-357
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    • 2005
  • This study evaluated the binding abilities of rabbit anti-ovalbumin (OVA) immunoglobulin G (IgG) and egg-allergic patient IgE on gamma-irradiated OVA during proteolysis using pepsin and trypsin. The concentrations of both the intact and the irradiated OVAs decreased during proteolysis when detected with IgG However, when detected by patient IgE the concentration of the intact OVA decreased up to 30 min after the trypsin treatment and increased thereafter. Irradiated OVA detected by patient IgE showed a lower initial concentration (0.16%) than that of the intact OVA, and this reduced concentration was maintained stably. The results indicate that irradiation, rather than enzymatic treatment, could reduce the binding of the irradiated and enzyme-treated OVA. Therefore, gamma irradiation has potential as an effective method to reduce OVA-induced allergy and may enhance the safety of egg-allergic individuals.

Content Comparison on Dietary Fiber and Rutin of Korean Buckwheat according to Growing District and Classification (한국산 메밀의 산지 및 종류에 따른 식이섬유와 루틴의 함량비교)

  • Lee, Mi-Sook;Sohn, Kyung-Hee
    • Korean journal of food and cookery science
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    • v.10 no.3
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    • pp.249-253
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    • 1994
  • In this study, total dietary fiber contents, acid detergent fiber contents lignin, water binding capacity (WBC), iron binding capacity of ADF and rutin contents from Korean buckwheats were determined. Total dietary fiber (TDF) content was 3.14% in kyungbuk rice buckwheat, 5.65% in kangwon hull buckwheat and 3.35% in kangwon rice buckwheat. Acid detergent fiber (ADF) content was 2.47, 4.46 and 2.94%, respectively and lignin content was 1.14, 1.60 and 1.08% respectively. The water binding capacity of these three sample dietary fiber showed that TDF were in the range of 2.87~3.88 g %H_2O$/g DF and ADF were in the range of 4.62~5.26 g %H_2O$/g DF. The iron binding capacity of ADF at pH 5.0, 6.0, 7.0 were 79.11, 78.01, 46.16% in kyungbuk rice buckwheat, 70.63, 63.83, 53.60% in kangwon hull buckwheat and 77.67, 76.33, 50.25% in kangwon rice buckwheat. The rutin contents of these samples and their hulls were 13.54~16.41 mg/100 g groats and 12.13~14.24 mg/100 g in hulls. The rutin content of kyungbuk rice buckwheat was the highest.

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Ethanol-induced Activiationof Transcription Factor NF-$\kappa$B and AP-1 in C6 Glial Cells

  • Park, Jae -Won;Shim, Young-Sup
    • Preventive Nutrition and Food Science
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    • v.4 no.3
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    • pp.209-214
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    • 1999
  • In this study, the effectof ethanol and acetaldehyde on DNA binding activities of NF-$textsc{k}$B and AP-1 were evaluated in C6 rat glial cells. Both NF-$textsc{k}$B and AP-1 are important transcription factors for the expression of various cytokines in glial cells. Our data showed that neither ethanol nor acetaldehyde induced conspicuous cell death of C6 cells at clinically realistic concentrations. When the DNA binding activities of nuclear NF-$textsc{k}$B and AP-1 were estimated using electrophoretic mobility shift assay (EMSA), ethanol(0.3%) or acetaldehyde(1mM) induced transient activation of these transcription factors, which attained peak levels at 4~8 hours and declined to basal levels at 12 hours after treatement . The supershift analysis showed that the increased activities of NF-$textsc{k}$B in ethanol/acetaldehyde-treated C6 cells were due to the preferential induction of p65/p50 heterodimer complex. The DNA binding activities of these transcriptional factors decreased below basal levels when cells were cultured with either ethanol or acetaldehyde for 24 hours, and showed the inhibitory effect of chronic ehtanol /acetaldehyde treatment on the activities of these transsriptional factors. Our data indicate that either ethanol or acetaldehyde can induce functional changes of glial cells throught bi-directional modulation of NF-$textsc{k}$B and AP-1 DNA binding activities.

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A Series of Quinoline-2-carboxylic Acid Derivatives: New Potent Glycine Site NMDA Receptor Antagonists

  • 김란희;최진일;최승원;이광숙;정영식;박우규;성철민;박노상
    • Bulletin of the Korean Chemical Society
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    • v.18 no.9
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    • pp.939-945
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    • 1997
  • Several types of 4-substituted-quinoline-2-carboxylic acid derivatives possessing different substituents at C4-position such as sulfonyl, phosphonyl, carbonyl groups, or a flexible alkyl chain have been synthesized and evaluated for their in vitro antagonistic activity at the glycine site on the N-methyl-D-aspartate (NMDA) receptor. Of them, 5,7-dichloro-4-(tolylsulfonylamino)-quinoline-2-carboxylic acid 9 was found to have the best in vitro binding affinity with IC50 of 0.57 μM. On the other hand, in compounds 21 and 22 the introduction of flexible alkyl chains on C4 of the quinoline mother nuclei caused a significant decrease of the in vitro binding affinity. In addition, replacement of polar carboxylic acid group on C2 by neutral bioisosteres in compounds 23a-d also seems to be disadvantageous to in vitro activity. In the structure-activity relationship (SAR) study of the 4-substituted quinoline-2-carboxylic acid acid derivatives, it was realized that the substitution pattern on C4 significantly influences on the binding affinity for the glycine site of NMDA receptor and the binding affinity might be increased by the introduction of a suitable electron rich substituent at C4 which has the ability of H-bonding donor.

Comparative Analysis of CCR2 and CCR5 Binding Sites to Facilitate the Development of Dual Antagonists: An in Silico Study

  • Kothandan, Gugan
    • Journal of Integrative Natural Science
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    • v.5 no.1
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    • pp.22-26
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    • 2012
  • Chemokine receptor antagonists have potential applications in field of drug discovery. Although the chemokine receptors are G-protein-coupled receptors, their cognate ligands are small proteins (8 to 12 kDa), and so inhibiting the ligand/receptor interaction has been challenging. In particular, CCR2 and CCR5 and their ligands have been implicated in the pathophysiology of a number of diseases, including rheumatoid arthritis and multiple sclerosis. Based on their roles in disease, they have been attractive targets for the pharmaceutical industry, targeting both CCR2 and CCR5 could be a useful strategy. Because of the importance of these receptors, providing information regarding the binding site is of prime importance. Herein, we report the comparison of CCR2 of CCR5 binding sites both sequentially as well as structurally. We also urged the importance of crucial residues in the binding site, to facilitate the development of dual antagonists targeting both the receptors. These results could also be useful for the design of novel and potent dual CCR2 and CCR5 antagonists using structure based drug design.

$Interferon-{\Upsilon}$ and Lipopolysaccaride Induce Mouse Guanylate-Binding Protein 3 (mGBP3) Expression in the Murine Macrophage Cell Line RAW264-7

  • Han, Byung-Hee
    • Archives of Pharmacal Research
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    • v.22 no.2
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    • pp.130-136
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    • 1999
  • Mouse guanylate-binding protein 3 (mGBP3) is a 71-kDa GTPase which belongs to GTP-binding protein family. The present study showed that the expression of mGBP3 transcript was readily induced in a dose dependent fashion in the macrophage cell line RAW264.7 treated with either $interferon-{\gamma} (IFN-\gamma)$ or lipopolysaccaride (LPS). The expression of mGBP3 protein was also apparent by 4 and 6 h after the treatment of cells with IFN-\gamma (100 U/ml) or LPS ($1{\mu}g/ml$) , and remained at palteau for at least 24 h. Cycloheximide ($10{\mu}g/ml$) had no effect on the $IFN-\gamma-$ or LPS-induced mGBP3 expression, suggesting that the mGBP3 induction did not require further protein synthesis. Interestingly, a protein kinase C (PKC) inhibitor staurosporine (50 nM) abolished the induction of mGBP3 expression by LPS, but not by $IFN-{\gamma}$. These findings suggest that mGBP3 may be involved in the macrophage activation process and both IFN-\gamma and LS induce the mGBP3 expression through distinct signal transduction pathways.

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Structural Insights into the Interaction of Terpenoids with Streptomyces avermitilis CYP107P2

  • Eunseo Jeong;Vitchan Kim;Changmin Kim;Yoo-bin Lee;Donghak Kim
    • Biomolecules & Therapeutics
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    • v.32 no.4
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    • pp.474-480
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    • 2024
  • Streptomyces avermitilis genome includes 33 genes encoding monooxygenation-catalyzing cytochrome P450 enzymes. We investigated the structure of CYP107P2 and its interactions with terpenoid compounds. The recombinant CYP107P2 protein was expressed in Escherichia coli and the purified enzyme exhibited a typical P450 spectrum upon CO-binding in its reduced state. Type-I substrate-binding spectral titrations were observed with various terpenoid compounds, including α-pinene, β-pinene, α-terpinyl acetate, and (+)-3-carene. The calculated binding affinities (Kd) ranged from 15.9 to 50.8 µM. The X-ray crystal structure of CYP107P2 was determined at 1.99 Å resolution, with a well-conserved overall P450 folding conformation. The terpenoid compound docking models illustrated that the structural interaction between monoterpenes and CYP107P2, with the distance between heme and terpenes ranging from 3.4 to 5.4 Å, indicates potential substrate binding for P450 enzyme. This study suggests that CYP107P2 is a Streptomyces P450 enzyme capable of catalyzing terpenes as substrates, signifying noteworthy advancements in comprehending a novel P450 enzyme's involvement in terpene reactions.