• Title/Summary/Keyword: Data Binding

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Secondary structure analysis of MRA1997 from Mycobacterium tuberculosis and characterization of DNA binding property

  • Kim, Hyo Jung;Lee, Ki-Young;Kim, Yena;Kwon, Ae-Ran;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.2
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    • pp.36-40
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    • 2016
  • MRA1997 is a highly conserved protein from mycobacterial strains. However, no structural and functional information is associated with it. Thus, to obtain details about structure and function of this protein, we have utilized NMR spectroscopy. The recombinant MRA1997 was highly purified and its DNA binding mode was characterized. The tertiary structure of MRA1997 was modeled on the basis of our NMR chemical shift data combined with the webserver CS23D. The binding of MRA1997 with DNA was first monitored by electrophoresis mobility shift assays. The residues involved in DNA binding are identified using NMR chemical shift perturbation experiments. Based on our study, we suggest that MRA1997 interacts with DNA and may play an important role in Mycobacterium tuberculosis physiology.

Differentially Expressed Genes in Marine Medaka Fish (Oryzias javanicus) Exposed to Cadmium

  • Woo, Seon-Ock;Son, Sung-Hee;Park, Hong-Seog;Vulpe, Chris D.;Ryu, Jae-Chun;Yum, Seung-Shic
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.293-299
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    • 2008
  • To screen the differentially expressed genes in cadmuim-exposed marine medaka fish (Oryzias javanicus), a candidate marine test fish for ecological toxicity, the differential display polymerase chain reaction (DD-PCR) was carried out, since the genome-wide gene expression data are not available in this fish species yet. A total of 35 clones were isolated from cadmium-exposed fish and their nucleotide sequences were analyzed. The differentially expressed gene candidates were categorized to response to stimulus (3); ion binding (3); DNA binding (1); protein binding (6); carbohydrate binding (1); metabolic process (4); biological regulation (3); cellular process (2); protein synthesis (2); catalytic activity (2); sense of sight (1); immune (1); neurohormone (1); signaling activity (1); electron carrier activity (1) and others (3). For real-time quantitative RT-PCR, we selected catalase, glucose-6-phosphate dehydrogenase, heat shock protein 70, and metallothionein and confirmed that cadmium exposure enhanced induction of these four genes.

Isolation of Proteins that Specifically Interact with the ATPase Domain of Mammalian ER Chaperone, BiP

  • Chung, Kyung-Tae;Lee, Tae-Ho;Kang, Gyong-Suk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.3
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    • pp.192-198
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    • 2003
  • BiP, immunoglobulin binding protein, is an ER homologue of Hsp70. However, unlit other Hsp70 proteins, regulatory protein(s) for BiP has not been identified. Here, we demo strafed the presence of potential regulatory proteins for BiP using a pull -down assay. Since BiP can bind any unfolded protein, only the ATPase domain of BiP was used for the pull -down assay in order to minimize nonspecific binding. The ATPase domain was cloned to produce recombinant protein, which was then conjugated to CNBr-activated agarose. The structural conformation and ATP hydrolysis activity of the recombinant ATPase domain were similar to those of the native protein, light proteins from metabolically labeled mouse plasmacytoma cells specifically bound to the recombinant ATPase protein. The binding of these proteins was inhibited by excess amounts of free ATPase protein, and was dependent on the presence of ATP. These proteins were eluted by ADP. Of these proteins, Grp170 and BiP where identified. while the other were not identified as known ER proteins, from Western blot analyses. The presence of the ATPase-binding proteins for BiP was first demonstrated in this study, and our data suggest similar regulatory machinery for BiP may exist in the ER, as found in prokaryotes and other cellular compartments.

Prediction of Mammalian MicroRNA Targets - Comparative Genomics Approach with Longer 3' UTR Databases

  • Nam, Seungyoon;Kim, Young-Kook;Kim, Pora;Kim, V. Narry;Shin, Seokmin;Lee, Sanghyuk
    • Genomics & Informatics
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    • v.3 no.3
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    • pp.53-62
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    • 2005
  • MicroRNAs play an important role in regulating gene expression, but their target identification is a difficult task due to their short length and imperfect complementarity. Burge and coworkers developed a program called TargetScan that allowed imperfect complementarity and established a procedure favoring targets with multiple binding sites conserved in multiple organisms. We improved their algorithm in two major aspects - (i) using well-defined UTR (untranslated region) database, (ii) examining the extent of conservation inside the 3' UTR specifically. Average length in our UTR database, based on the ECgene annotation, is more than twice longer than the Ensembl. Then, TargetScan was used to identify putative binding sites. The extent of conservation varies significantly inside the 3' UTR. We used the 'tight' tracks in the UCSC genome browser to select the conserved binding sites in multiple species. By combining the longer 3' UTR data, TargetScan, and tightly conserved blocks of genomic DNA, we identified 107 putative target genes with multiple binding sites conserved in multiple species, of which 85 putative targets are novel.

Rat Liver $AT_1$ Receptor Binding Analysis for Drug Screening

  • Lee, Sunghou;Lee, Buyean;Hwasup Shin;Jaeyang Kong
    • Biomolecules & Therapeutics
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    • v.3 no.1
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    • pp.21-27
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    • 1995
  • The only compounds with antagonistic activity via AT$_1$receptor, one of two subtypes of angiotensin II (AII) receptor, have been demonstrated to block the vasoconstriction effects of AII and thereby provide therapeutic potential. This initiated the search for compounds with high specific affinity to AT$_1$receptor and their effective screening methods. The radioligand binding assay for the AII receptor is regarded as the primary method for the evaluation of AT$_1$receptor antagonists for their activity. In this paper, we characterized the liver AT$_1$receptor and describe the efficient method of the radioligand binding assay using rat liver as a source of AT$_1$receptor. Equilibrium binding studies with rat adrenal cortex, adrenal medulla, liver and bovine adrenal showed that the specific bindings of [$^3$H] AII were saturable in all tissues and the Scatchard plots of those data were linear, suggesting a single population of binding sites. Hill slopes were very near to the unity in all tissues. Kinetic studies of [$^3$H) AII binding in rat liver homogenates yielded two association rate constants, 4.10$\times$10$^{7}$ M$^{-1}$ min$^{-1}$ and 4.02$\times$10$^{9}$ M$^{-1}$ min$^{-1}$ , with a single dissociation rate constant, 7.07$\times$10$^{-3}$ min-$^{-1}$ , possibly due to the partial dissociation phenomenon. The rank order of inhibition potencies of [$^3$H] AII binding in rat liver was AII>Sarile>Losartan>PD 123177. Rat liver homogenates revealed to have very high density of homogeneous population of the AT$_1$receptor subtype, as the specifically bound [$^3$H] AII was not inhibited by PD 123177, the nonpeptide antagonist of AT$_2$. The results of this study demonstrated that the liver homogenates from rats could be the best receptor preparation for the AT$_1$receptor binding assay and provide an efficient system for the screening of newly synthesized candidate compounds of AT$_1$receptor antagonist.

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A Point Mutation at the C-Terminal Half of the Repressor of Temperate Mycobacteriophage L1 Affects Its Binding to the Operator DNA

  • Ganguly, Tridib;Chattoraj, Partho;Das, Malabika;Chanda, Palas K.;Mandal, Nitai.C.;Lee, Chia Y.;Sau, Subrata
    • BMB Reports
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    • v.37 no.6
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    • pp.709-714
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    • 2004
  • The wild-type repressor CI of temperate mycobacteriophage L1 and the temperature-sensitive (ts) repressor CIts391 of a mutant L1 phage, L1cIts391, have been separately overexpressed in E. coli. Both these repressors were observed to specifically bind with the same cognate operator DNA. The operator-binding activity of CIts391 was shown to differ significantly than that of the CI at 32 to $42^{\circ}C$. While 40-95% operator-binding activity was shown to be retained at 35 to $42^{\circ}C$ in CI, more than 75% operator-binding activity was lost in CIts391 at 35 to $38^{\circ}C$, although the latter showed only 10% less binding compared to that of the former at $32^{\circ}C$. The CIts391 showed almost no binding at $42^{\circ}C$. An in vivo study showed that the CI repressor inhibited the growth of a clear plaque former mutant of the L1 phage more strongly than that of the CIts391 repressor at both 32 and $42^{\circ}C$. The half-life of the CIts391-operator complex was found to be about 8 times less than that of the CI-operator complex at $32^{\circ}C$. Interestingly, the repressor-operator complexes preformed at $0^{\circ}C$ have shown varying degrees of resistance to dissociation at the temperatures which inhibit the formation of these complexes are inhibited. The CI repressor, but not that of CIts391, regains most of the DNA-binding activity on cooling to $32^{\circ}C$ after preincubation at 42 to $52^{\circ}C$. All these data suggest that the 131st proline residue at the C-terminal half of CI, which changed to leucine in the CIts391, plays a crucial role in binding the L1 repressor to the cognate operator DNA, although the helix-turn-helix DNA-binding motif of the L1 repressor is located at its N-terminal end.

GTPase Activity Analysis of eRF3 in Euplotes octocarinatus

  • Song, Li;Dong, Jun-Li;Zhao, Ya-Qin;Chai, Bao-Feng;Liang, Ai-Hua
    • Journal of Microbiology and Biotechnology
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    • v.20 no.9
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    • pp.1283-1287
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    • 2010
  • In eukaryotes, eRF3 participates in translation termination and belongs to the superfamily of GTPases. In this work, the dissociation constants for nucleosides bound to Euplotes octocarinatus eRF3 in the presence and absence of eRF1a were determined using fluorescence spectra methods. Furthermore, a GTP hydrolyzing assay of eRF3 was carried out using an HPLC method, and the kinetic parameters for GTP hydrolysis by eRF3 were determined. Consistent with data from humans, the results showed that eRF1a promoted the binding of GTP to eRF3 and the GTP hydrolyzing activity of eRF3. However, in contrast to the lack of GTP binding in the absence of eRF1 in human eRF3, the E. octocarinatus eRF3 was able to bind GTP by itself. The nucleotide binding affinity of the E. octocarinatus eRF3 also differed from the human data. A structure model and amino acid sequence alignment of potential G domains indicated that these differences may be due to valine 317 and glutamate 452 displacing the conserved glycine and lysine involved in GTP binding.

A Development of DCS Binding Delay Analysis System based on PC/Ethernet and Realtime Database

  • Gwak, Kwi-Yil;Lee, Sung-Woo;Lim, Yong-Hun;Lee, Beom-Seok;Hyun, Duck-Hwa
    • 제어로봇시스템학회:학술대회논문집
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    • 2005.06a
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    • pp.1571-1576
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    • 2005
  • DCS has many processing components and various communication elements. And its communication delay characteristic is affected diverse operating situation and context. Especially, binding signal which traversed from one control-node to another control-node undergo all sort of delay conditions. So its delay value has large deviation with the lapse of time, and the measurement of delay statistics during long time is very difficult by using general oscilloscope or other normal instruments. This thesis introduces the design and implementation of PC-based BDAS(Binding Delay Analysis System) System developed to overcomes these hardships. The system has signal-generator, IO-card, data-acquisition module, delay-calculation and analyzer module, those are implemented on industrial standard PC/Ethernet hardware and Windows/Linux platforms. This system can detect accurate whole-system-wide delay time including io, control processing and network delay, in the resolution of msec unit, and can analyze each channel's delay-historic data which is maintained by realtime database. So, this system has strong points of open system architecture, for example, user-friendly environment, low cost, high compatibility, simplicity of maintenance and high extension ability. Of all things, the measuring capability of long-time delay-statistics obtained through historic-DB make the system more valuable and useful, which function is essential to analyze accurate delay performance of DCS system. Using this system, the verification of delay performance of DCS for nuclear power plants is succeeded in KNICS(Korea Nuclear Instrumentation & Control System) projects

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Investigating Binding Area of Protein Surface using MCL Algorithm (MCL 알고리즘을 이용한 단백질 표면의 바인딩 영역 분석 기법)

  • Jung, Kwang-Su;Yu, Ki-Jin;Chung, Yong-Je;Ryu, Keun-Ho
    • The KIPS Transactions:PartD
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    • v.14D no.7
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    • pp.743-752
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    • 2007
  • Proteins combine with other materials to achieve their function and have similar function if their active sites are similar. Thus we can infer the function of protein by identifying the binding area of proteins. This paper suggests the novel method to select binding area of protein using MCL (Markov Cluster) algorithm. We construct the distance matrix from surface residues distance on protein. Then this distance matrix is transformed to connectivity matrix for applying MCL process. We adopted Catalytic Site Atlas (CSA) data to evaluate the proposed method. In the experimental result using CSA data (94 selected single chain proteins), our algorithm detects the 91 (97%) binding area near by active site of each protein. We introduced a new geometrical features and this mainly contributes to reduce the time to analyze the protein by selecting the residues near by active site.

Electronic properties of monolayer silicon carbide nanoribbons using tight-binding approach

  • Chuan, M.W.;Wong, Y.B.;Hamzah, A.;Alias, N.E.;Sultan, S. Mohamed;Lim, C.S.;Tan, M.L.P.
    • Advances in nano research
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    • v.12 no.2
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    • pp.213-221
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    • 2022
  • Silicon carbide (SiC) is a binary carbon-silicon compound. In its two-dimensional form, monolayer SiC is composed of a monolayer carbon and silicon atoms constructed as a honeycomb lattice. SiC has recently been receiving increasing attention from researchers owing to its intriguing electronic properties. In this present work, SiC nanoribbons (SiCNRs) are modelled and simulated to obtain accurate electronic properties, which can further guide fabrication processes, through bandgap engineering. The primary objective of this work is to obtain the electronic properties of monolayer SiCNRs by applying numerical computation methods using nearest-neighbour tight-binding models. Hamiltonian operator discretization and approximation of plane wave are assumed for the models and simulation by applying the basis function. The computed electronic properties include the band structures and density of states of monolayer SiCNRs of varying width. Furthermore, the properties are compared with those of graphene nanoribbons. The bandgap of ASiCNR as a function of width are also benchmarked with published DFT-GW and DFT-GGA data. Our nearest neighbour tight-binding (NNTB) model predicted data closer to the calculations based on the standard DFT-GGA and underestimated the bandgap values projected from DFT-GW, which takes in account the exchange-correlation energy of many-body effects.