• 제목/요약/키워드: binding number

검색결과 530건 처리시간 0.028초

ErbB3 binding protein 1 contributes to adult hippocampal neurogenesis by modulating Bmp4 and Ascl1 signaling

  • Youngkwan Kim;Hyo Rim Ko;Inwoo Hwang;Jee-Yin Ahn
    • BMB Reports
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    • 제57권4호
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    • pp.182-187
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    • 2024
  • Neural stem cells (NSCs) in the adult hippocampus divide infrequently; the endogenous molecules modulating adult hippocampal neurogenesis (AHN) remain largely unknown. Here, we show that ErbB3 binding protein 1 (Ebp1), which plays important roles in embryonic neurodevelopment, acts as an essential modulator of adult neurogenic factors. In vivo analysis of Ebp1 neuron depletion mice showed impaired AHN with a low number of hippocampal NSCs and neuroblasts. Ebp1 leads to transcriptional repression of Bmp4 and suppression of Ascl1 promoter methylation in the dentate gyrus of the adult hippocampus reflecting an unusually high level of Bmp4 and low Ascl1 level in neurons of Ebp1-deficient mice. Therefore, our findings suggests that Ebp1 could act as an endogenous modulator of the interplay between Bmp4 and Ascl1/Notch signaling, contributing to AHN.

개 정자의 수정능력 검정을 위한 Hamster test의 이용 가능성 (Availability of Hamster Test to Assess the Fertilizing Capacity of Dog Sperm)

  • 이해이;김용준
    • 한국임상수의학회지
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    • 제10권1호
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    • pp.1-10
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    • 1993
  • To investigate the availability of hamster test in assaying the fertilizing capacity of dog sperm and the effect of canine sperm motility on sperm binding and penetration, semen were collected from four dogs(three dogs had been proven to be fertile and one dog to be subfertile during the past two years) and then preserved in BWW(Biggers, Whitten, Whittingham) medium for about 20 hours. The semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding) and ova penetrated by sperm(penetration), and also numbers of both bound and penetrated sperm per ovum. In comparison between fertile dogs and a subfertile dog, the rate of sperm binding was higher in fertile dogs than the subfertile dog(p<0.01, p<0.05). The number of bound sperm per ovum was considerably higher in a fertile dog than the subfertile dog((p<0.01), and also difference of number of the bound sperm was obtained among the fertile dogs(p<0.01, p<0.05). The rate of penetration as well as the number of penetrated sperm per ovum was higher in the fertile dogs than the subfertile dog(p<0.01, p<0.05). In fertile dogs. the canine semen preserved at 4$^{\circ}C$ for 18 to 22 hours showed from 30 to 80% motility at Insemination, however, no difference in hamster test was obtained according to different degree of sperm motility. These results indicated that hamster test would be of avail in assaying the fertilizing capacity of dog sperm.

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Bacterial Overexpression and Denaturing Purification of VPS34-Binding Domain of Beclin 1

  • Baek, Jong-Hyuk;Jung, Juneyoung;Seo, Jeongbin;Kim, Jeong Hee;Kim, Joungmok
    • Journal of Microbiology and Biotechnology
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    • 제26권10호
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    • pp.1808-1816
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    • 2016
  • As a scaffolding subunit of the PIK3C3/VPS34 complex, Beclin 1 recruits a variety of proteins to class III phosphatidylinositol-3-kinase (VPS34), resulting in the formation of a distinct PIK3C3/VPS34 complex with a specific function. Therefore, the investigation of a number of Beclin 1 domains required for the protein-protein interactions will provide important clues to understand the PIK3C3/VPS34 complex, of which Beclin1-VPS34 interaction is the core unit. In the present study, we have designed a bacterial overexpression system for the Beclin 1 domain corresponding to VPS34 binding (Vps34-BD) and set up the denaturing purification protocol due to the massive aggregation of Vps34-BD in Escherichia coli. The expression and purification conditions determined in this study successfully provided soluble and functional Vps34-BD.

Biochemical characterization of the lipid-binding properties of a broccoli cuticular wax-associated protein, WAX9D, and its application

  • Ahn, Sun-Young;Kim, Jong-Min;Pyee, Jae-Ho;Park, Heon-Yong
    • BMB Reports
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    • 제42권6호
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    • pp.367-372
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    • 2009
  • In this study, we showed that WAX9D, a nonspecific lipid-transfer protein found in broccoli, binds palmitate (C16) and stearate (C18) with dissociation constants of 0.56 ${\mu}M$ and 0.52 ${\mu}M$, respectively. WAX9D was fused to thioredoxin protein by genetic manipulation to enhance its solubility. The data revealed strong interaction of Trx-WAX9D with palmitate and stearate. The dissociation constants of Trx-WAX9D for palmitate and stearate were 1.1 ${\mu}M$ and 6.4 ${\mu}M$, respectively. The calculated number of binding sites for palmitate and stearate was 2.5 to 2.7, indicating that Trx-WAX9D can bind three molecules of fatty acids. Additionally, Trx-WAX9D was shown to inhibit the apoptotic effect of palmitate in endothelial cells. Our data using Trx-WAX9D provide insight into the broad spectrum of its biological applications with specific palmitate binding.

Inhibitory Effects of Bovine Serum Albumin on Cytotoxicity and Mutagenicity of 6-Sulfooxymethylbenzo[a]pyrene

  • Cho, Young-Sik;Cho, Kyung-Joo;Chung, An-Sik
    • Toxicological Research
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    • 제16권3호
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    • pp.221-227
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    • 2000
  • A 6-sulfooxymethylbenzo[a]pyrene (SMBP), the ultimate metabolite of methyl-substituted benzo[a]pyrene (BP), has been found to be carcinogenic in mice. These properties may be attributable to its strong reactivity with cellular macromolecules such as DNA. However, serum and its major constituent albumin attenuated significantly the cytotoxicity and mutagenicity of 5MBP in bacterial and mammalian cell systems. This inhibitory activity of serum against 5MBP-induced cytotoxicity and mutagenicity in Chinese hamster V79 cells appears to be caused by the reduced macromolecular adducts such as DNA and proteins, but serum failed to reduce 5MBP binding to naked calf thymus DNA. A number of proteins in the serum could act as nucleophiles that are able to intercept reactive chemicals through covalent binding. Albumin present in the plasma seems to be one of major components responsible for direct binding with 5MBp, thereby reducing its reactivity to genetic materials. We here determined which fraction is preferential for 5MBP binding through fractionation of 5MBP-treated serum with ammonium sulfate. The albumin-containing fraction had slightly more affinity for 5MBP than the immunoglobulin-containing fraction. Our results indicate that the covalent modification of plasma proteins may reduce 5MBP-induced damage.

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B3(Fab)-streptavidin Tetramer Has Higher Binding Avidity than B3(scFv)-streptavidin Tetramer

  • Won, Jae-Seon;Kang, Hye-Won;Nam, Pil-Won;Choe, Mu-Hyeon
    • Bulletin of the Korean Chemical Society
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    • 제30권5호
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    • pp.1101-1106
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    • 2009
  • Multivalent and multi-specific antibodies can provide valuable tools for bio-medical research, diagnosis and therapy. In antigen-antibody interactions, the avidity of antibodies depends on the affinity and the number of binding sites.$^1$ As artificial multivalent antibody agents, single chain Fv-streptavidin fusion tetramer proteins $(scFv-SA)_4$ have been previously tested.$^{1,\;2}$ Although, the Fab domain is known to be more stable than scFv in animal models,$^{3,\;4}$ it has never been used to make a multivalent agent with a streptavidin fusion. In this study, we prepared tetra-valent $(Fab-cSA)_4$ by fusing Fab with core streptavidin (cSA). This molecule was made using inclusion body production, refolding and chromatography purification. Affinities of the Fab-cSA tetramer and a scFv-cSA tetramer to a cell surface antigen were compared by ELISA using biotin-HRP. The Fab-cSA tetramer showed higher binding avidity than the scFv-cSA tetramer. The higher binding avidity of the Fab-cSA tetramer demonstrates its potential as a therapeutic agent for target-specific antibody therapy.

Sorption Behavior of 1-Methylcyclopropene on Adsorbing Agents for Use in Extending the Freshness of Postharvest Food Products

  • Lee, Youn-Suk;Shin, Han-Seung
    • Food Science and Biotechnology
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    • 제15권4호
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    • pp.572-577
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    • 2006
  • The physiochemical interactions of 1-methylcyclopropene (1-MCP) and adsorbing agents can be described using a very powerful tool, inverse gas chromatography (IGC). Sorption behavior of 1-MCP on various adsorbing agents was assessed using the profile peaks of 1-MCP at an infinite dilution concentration using the IGC technique. Chromatogram peaks of 1-MCP adsorption were not observed for the adsorbing agent activated carbon. The forms of sorption isotherms followed Henry's law, and behaved according to the binding site theory. Specific retention volume and distribution coefficients for 1-MCP on the adsorbing agents were determined at 50, 60, 70, and $80^{\circ}C$, respectively. Silica gel had a much higher number of binding sites for 1-MCP compared to Tenax-TA and activated clay agents. Meanwhile, activated carbon proved to be a very strong binding agent for 1-MCP based on 1-MCP efficiency experiments on the selected adsorbing agents. However, as a proper means of delivering 1-MCP molecules to fresh food products, activated carbon is not fit for the binding and release of 1-MCP gas under dry or high humidity conditions because activated carbon has a strong affinity for 1-MCP, even when treated with distilled water.

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

  • Kothandan, Gugan
    • 통합자연과학논문집
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    • 제5권1호
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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.

Redirect 공격과 DoS 공격에 안전한 MIPv6 바인딩 업데이트 프로토콜 (MIPv6 Binding Update Protocol Secure Against both Redirect and DoS Attacks)

  • 강현선;박창섭
    • 정보보호학회논문지
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    • 제15권5호
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    • pp.115-124
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    • 2005
  • 본 논문에서는 모바일 노드(mobile node, MN)와 대응노드(correspondent node, CN)사이의 바인딩 업데이트 (binding update, BU) 프로토콜에 대해 기존 프로토콜들에서 알려진 Redirect 공격과 DOS(denial-of service) 공격을 방지하고 효율성을 향상시키기 위해 새로운 BU 프로토콜을 제안한다. 제안 프로토콜에서 홈 에이전트(home agent, HA)는 BU 메시지의 유효성을 확인하는 인증서버(authentication server)의 기능과 MN과 CN를 위한 세션키 분배센터의 기능을 수행한다. 또한 CGA(cryptographically generated address)를 기반으로 한 stateless Diffie-Hellman 키 합의 기법을 소개하고, 제안 프로토콜의 안전성을 기존 프로토콜과 함께 비교하고 분석한다. 제안 프로토콜은 메시지 수, 계산적 오버헤드 측면에서 기존기법보다 효율적이며, Redirect와 DoS 공격으로부터 안전하다.

Thermodynamic and Structural Studies on the Human Serum Albumin in the Presence of a Polyoxometalate

  • Ajloo, D.;Behnam, H.;Saboury, A.A.;Mohamadi-Zonoz, F.;Ranjbar, B.;Moosavi-Movahedi, A.A.;Hasani, Z.;Alizadeh, K.;Gharanfoli, M.;Amani, M.
    • Bulletin of the Korean Chemical Society
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    • 제28권5호
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    • pp.730-736
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    • 2007
  • The interaction of a polyoxometal (POM), K6SiW11Co(H2O)O39.10H2O (K6) as a Keggin, with human serum albumin (HSA) was studied by different methods and techniques. Binding studies show two sets of binding sites for interaction of POM to HSA. Binding analysis and isothermal calorimetery revealed that, the first set of binding site has lower number of bound ligand per mole of protein (ν), lower Hill constant (n), higher binding constant (K), more negative entropy (ΔS) and more electrostatic interaction in comparison to the second set of binding site. In addition, differential scanning calorimetery (DSC) and spectrophotometery data showed that, there are two energetic domains. The first domain is less stable (lower Tm and Cp) which corresponds to the tail segment of HSA and another with more stability is related to the head segment of HSA. Polyoxometal also decreases the stability of protein as Tm, secondary and tertiary structure as well as quenching of the fluorescence decrease. On other hand, perturbations in tertiary structure are more than secondary structure.