• 제목/요약/키워드: Binding Mechanism

검색결과 985건 처리시간 0.025초

Characterization of binding specificity using GST-conjugated mutant huntingtin epitopes in surface plasmon resonance (SPR)

  • Cho, Hang-Hee;Kim, Tae Hoon;Kim, Hong-Duck;Cho, Jae-Hyeon
    • 한국동물위생학회지
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    • 제44권4호
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    • pp.185-194
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    • 2021
  • Polyglutamine extension in the coding sequence of mutant huntingtin causes neuronal degeneration associated with the formation of insoluble polyglutamine aggregates in Huntington's disease (HD). Mutant huntingtin can form aggregates within the nucleus and processes of neurons possibly due to misfolding of the proteins. To better understand the mechanism by which an elongated polyglutamine causes aggregates, we have developed an in vitro binding assay system of polyglutamine tract from truncated huntingtin. We made GST-HD exon1 fusion proteins which have expanded polyglutamine epitopes (e.g., 17, 23, 32, 46, 60, 78, 81, and 94 CAG repeats). In the present emergence of new study adjusted nanotechnology on protein chip such as surface plasmon resonance strategy which used to determine the substance which protein binds in drug discovery platform is worth to understand better neurodegenerative diseases (i.e., Alzheimer disease, Parkinson disease and Huntington disease) and its pathogenesis along with development of therapeutic measures. Hence, we used strengths of surface plasmon resonance (SPR) technology which is enabled to examine binding specificity and explore targeted molecular epitope using its electron charged wave pattern in HD pathogenesis utilize conjugated mutant epitope of HD protein and its interaction whether wild type GST-HD interacts with mutant GST-HD with maximum binding affinity at pH 6.85. We found that the maximum binding affinity of GST-HD17 with GST-HD81 was higher than the binding affinities of GST-HD17 with other mutant GST-HD constructs. Furthermore, our finding illustrated that the mutant form of GST-HD60 showed a stronger binding to GST-HD23 or GST-HD17 than GST-HD60 or GST-HD81. These results indicate that the binding affinity of mutant huntingtin does not correlate with the length of polyglutamine. It suggests that the aggregation of an expanded polyglutamine might have easily occurred in the presence of wild type form of huntingtin.

인삼 지용성성분인 유전독성억제효과와 작용기전 (Antigenotoxicity of Ginseng Petroleum Ether Extract and its Action Mechanism)

  • 허문영
    • 한국식품위생안전성학회지
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    • 제13권3호
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    • pp.243-251
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    • 1998
  • 백삼(Pampx ginseng C.A. Meyer, 4년근)을 가루로 한 다음 petroleum ether로 24시간 추출 후 감압 농축시켜 인삼 석유에텔추출물(GPE)로 하였다. 이때 GPE의 수율은 평균 2.24%였다. 또한 GPE를 petroleum ether 소량에 녹이고 petroleum ether:ether 용매계열로 silicagel column chromatography에 의한 partition을 실시하여 P1, P2, P3 , P4, P5의 5개의 분획을 얻었다. 본 실험에서는 소핵생성 물질인 N-methyl-N-nitrosourea(MNU) 및 benzo(a) pyrene[B(a)P]를 이용하여 소핵생성억제실험을 하였따. 상기의 5가지 분획 중에서 MNU에 대해서는 GPE, P1, P2에서 억제효과가 나타났다. 한편, B(a)P에 대해서는 GPE, P1, P2, P3 , P4, P5 모두에게 억제효과를 나타내었다. 그러나 여러 가지 활성을 보인 분획 중에서 여러 가지 양성대조물질들에 대해서 가장 두드러져 보이는 것은 P2였다. P2는 MNU, B(a)P 모두에 대해서 GPE와 유사한 억제경향을 보이면서 억제활성은 크게 나타난T다. EK라서 P2가 GPE가 보이는 소핵생성억제효과에 깊은 관련이 있는 물질로 판단되었다. 이와 같은 유전독성억제효과의 작용기전을 알기 위하여 B(a)P의 대사에 미치는 영향을 연구한 결과 GPE와 P2는 S-9 mix에 의해 유도된 B(a)P 대사를 억제시켰으며, DNA-B(a)P binding도 감소시켰다. 한편, MNU에 의한 DNA binding 과 O6-methyl guanine 및 7-methyl guanine 생성에 있어서도 억제적인 경향을 나타내었다. 따라서 GPE와 P2는 B(a)P와 같은 2차 발암 물질과 MNU와 같은 알킬화제에 의한 유전독성을 활성대사억제 및 methylation 억제 , DNA binding 억제 등의 기전으로 감소시키고 있는 것으로 판단되었다. 한편, 소핵생성억제 효과에서 가장 활성이 좋았던 P2를 NMR과 GC/MS결과, aliphatic ketone류의 혼합물로서 주성분은 분질량 330과 386의 두 개 물질이 함유되어 있는 혼합물이었으며 향후 계속적으로 분리 동정이 필요한 물질이다.

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Presence of A Negative Light Regulatory Factors Binding to the cab3 Promoter of Arabidopsis Thaliana

  • Kang, Ku-Seong
    • Journal of Photoscience
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    • 제5권4호
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    • pp.149-152
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    • 1998
  • Expression of light-harvesting chlorophyll a/b-binding protein gene(cab) is repressed in the dark and activited by light. However, the detail of its regulatory mechanism is not characterized so far. To identify the interactions of cis-acting elements and trans-acting factors involvedin this regulation, nuclear extracts from the light-grown and dark-adapted Arabidopsis thaliana leaves were anlayzed for mobility shift assay against 134bp fragments had two retarded bands and one retardation band, respectively, both in light-grown and dark-adapted bands in the dark-adapted tissues. A new retardation the cab 3 expression in the dark. Several light regulatory motifs are scattered in the 146 bp region of cab 3 promoter. One of the light-regulatory motifs could be the binding site for the negative regulatory factor.

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Ginsenoside Rg$_3$ inhibits NMDA receptors in rat cultured hippocampal neurons: possible involvement of a glycine-binding site

  • Rhim, Hye-Whon
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 2004년도 추계 학술대회 및 정기총회
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    • pp.7-11
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    • 2004
  • We previously reported that ginseng inhibited NMDA receptors in cultured hippocampal neurons. Here, we further examined the detailed mechanism of ginseng-mediated inhibition using its main active ingredient, ginsenoside Rg$_3$. Co-application of ginsenoside Rg$_3$ with increasing concentrations of NMDA did not change the EC$_{50}$ of NMDA to the receptor, suggesting ginsenoside Rg$_3$ inhibits NMDA receptors without competing with the NMDA-binding site. Ginsenoside Rg$_3$-mediated inhibition also occurred in a distinctive manner from the well-characterized NMDA receptor open channel blocker, MK-801, However, ginsenoside Rg$_3$ produced its effect in a glycine concentration-dependent manner and shifted the glycine concentration-response curve to the right without changing the maximal response, suggesting the role of ginsenoside Rg$_3$ as a competitive NMDA receptor antagonist. We also demonstrated that ginsenoside Rg$_3$ significantly protected neurons against NMDA insults. Therefore, these results suggest that ginsenoside Rg$_3$ protects NMDA-induced neuronal death via a competitive interaction with the glycine-binding site of NMDA receptors in cultured hippocampal neurons.

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뉴톤법에 의한 최적전력 조류계산 (Optimal Power Flow Study by The Newton's Method)

  • 황갑주
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 1989년도 하계종합학술대회 논문집
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    • pp.173-178
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    • 1989
  • Optimal Power Flow (OPF) solution by the Newton's method provides a reliable and robust method to classical OPF problems. The major challenge in algorithm development is to identify the binding inequalities efficiently. This paper propose a simple strategy to identify the binding set. From the mechanism of penalty shifting with soft penalty in trial iteration, a active binding sit is identified automatically. This paper also suggests a technique to solve the linear system whore coefficients are presented by the matrix. This implementation is highly efficient for sparsity programming. Case study for 3,5,14,118,190 bus and practrical KEPCO 305 bus system are performed as well.

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Yeast Two Hybrid Assay를 이용한 Lipocortin-1 결합 단백질 유전자의 분리 (Isolation of the Gene for Lipocortin-1 Binding Protein Using Yeast Two Hybrid Assay)

  • 이경화;김정우
    • 자연과학논문집
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    • 제9권1호
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    • pp.25-29
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    • 1997
  • Glucocorticoid에 의한 항염증 작용의 second messenger로 생각되어지는 annexin superfamily중 하나인 37 kDa의 단백질, lipocortin-1의 작용기작을 이해할 목적으로 in vivo에서 protein-protein interaction을 인식하는 yeast-based genetic assay인 yeast two assay를 통하여 lipocortin-1과 결합하는 단백질 유전자를 분리하여 조사하였다. 이 방법으로 실험을 수행한 결과 분리된 유전자가 human serine proteinase 유전자와 homology가 있는 것으로 밝혀졌다.

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The regulation of stress induced genes by yeast transcription factor GCN4

  • Seong Kimoon;Lee Jae Yung;Kim Joon
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2002년도 추계학술대회
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    • pp.135-139
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    • 2002
  • Yeast cells respond to condition of amino acid starvation by synthesizing GCN4, a typical eukaryotic transcriptional activator, which regulates the expression of many amino acids biosynthetic genes. By introducing point mutations in the DNA binding domain of GCN4, mutants with normal DNA binding activity but defective in transcriptional activity were isolated to identify unknown proteins that could suppress the mutant phenotype under an amino acid depletion condition. As a result, SSB(Stress-Seventy B) subfamily proteins were identified as suppressors of mutant GCN4. SSB proteins were known as a member of yeast hsp70 family that probably aids passage of nascent chain through ribosomes. Among them, the mechanism of suppression by SSB2 on the defective GCN4 mutant strains is under investigation. Gcn4p directly interacts with Ssb2p through the basic DNA binding domain of GCN4. It suggests the possibility that physical interaction might induce the transcriptional activation of Gcn4p.

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뉴톤법에 의한 최적전력 조류계산의 개선 (Optimal Power Flow Study by The Newton's Method)

  • Hwang, Kab-Ju
    • 대한전기학회논문지
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    • 제39권3호
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    • pp.223-231
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    • 1990
  • Optimal Power Flow (OPF) solution by the Newton's method provides a reliable and robust method to classical OPF problems. The major challenge in algorithm development is to identify the binding inequalities efficiently. This paper proposes a simple strategy to identify the binding set. From the mechanism of penalty shifting with soft penalty in trial iteration, an active binding set is identidied automatically. This paper also suggests a technique to solve the linear system whose coefficients are presented in the matrix from. This implementation is highly efficient for sparsity programming. Case studies for 3, 5, 14, 118 bus and practical TPC-190, KEPCO-306 bus systems are performed as well.

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Quantum chemical investigations on bis(bora)calix[4]arene: a fluorescent fluoride sensor

  • Jin, Jae Hyeok;Lee, Yoon Sup
    • EDISON SW 활용 경진대회 논문집
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    • 제2회(2013년)
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    • pp.77-88
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    • 2013
  • The computational study on the fluoride ion binding with bis(bora)calix[4]arene has been performed using density functional theory and ONIOM model. The computed structure and fluorescent behavior of bis(bora)calix[4]arene was corresponded to experiment value. The binding energy for fluoride anion is computed to be 28.05kJ/mol in the chloroform solution. We also predicted that this sensing mechanism is only valid for fluoride ion in halogens. By analyzing molecular orbitals, binding with fluoride ion reduces energy differences between HOMO and LUMO, which leads to fluorescent sensing.

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A Structure-Based Activation Model of Phenol-Receptor Protein Interactions

  • 이경희
    • Bulletin of the Korean Chemical Society
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    • 제18권1호
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    • pp.18-23
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    • 1997
  • Data from structure/activity studies in vir gene induction system have led to evaluate the working hypothesis of interaction between phenolic inducers and phenol binding proteins. The primary specificity in the association of a phenolic inducer with its receptor in our system is hypothesized to be the hydrogen bonding interactions through the ortho methoxy substituents as well as the proton transfer between the inducer and the binding protein. In this paper the proposed working model for phenol-mediating signal transduction was evaluated in several ways. The importance of the general acid-base catalysis was first addressed by the presence of an acidic residue and a basic residue in the phenol binding protein. Series of compounds were tested for vir gene expression activity to confirm the generation of a strong nucleophile by an acidic residue and an involvement of a basic residue as a proton acceptor. An attempt was made to correlate the pKa values of the phenolic compounds with vir gene induction activities as inducers to further support the proposed proton transfer mechanism. Finally, it was also observed that the regioselectively attached methoxy group on phenol compounds is required as the proper hydrogen bond acceptor.