• Title/Summary/Keyword: NS2A

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Selection and Target-Site Mapping of Peptides Inhibiting HCV NS5B Polymerase Using Phage Display

  • Kim, Min-Soo;Park, Chan-Hee;Lee, Jong-Ho;Myung, Hee-Joon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.328-333
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    • 2008
  • A series of pep tides binding to the HCV NS5B polymerase was selected from phage display peptide libraries. A conserved motif of Ser-Arg-X-Arg/Leu was identified among the selected peptides, and Pep2 (Trp-Ser-Arg-Pro-Arg-Ser-Leu) was chosen for further characterization. The binding of Pep2 to HCV NS5B in vivo was shown by a yeast two-hybrid assay and by subcellular colocalization analysis using immunofluorescence confocal microscopy. The in vitro interaction was also confirmed by GST pulldown assay. The replication of the HCV 1b subgenomic replicon was efficiently inhibited by the presence of the peptide. By using a subtractive biopanning against Pep2, the binding site of the peptide was mapped at the pocket of Pro388 to Pro391 in the thumb subdomain of the polymerase. A yeast two-hybrid analysis using Pro388Ala and Pro391Ala mutants of NS5B confirmed the binding.

Nonstructural Protein 5B of Hepatitis C Virus

  • Lee, Jong-Ho;Nam, In Young;Myung, Heejoon
    • Molecules and Cells
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    • v.21 no.3
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    • pp.330-336
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    • 2006
  • Since its identification in 1989, hepatitis C virus has been the subject of extensive research. The biology of the virus and the development of antiviral drugs are closely related. The RNA polymerase activity of nonstructural protein 5B was first demonstrated in 1996. NS5B is believed to localize to the perinuclear region, forming a replicase complex with other viral proteins. It has a typical polymerase structure with thumb, palm, and finger domains encircling the active site. A de novo replication initiation mechanism has been suggested. To date, many small molecule inhibitors are known including nucleoside analogues, non-nucleoside analogues, and pyrophosphate mimics. NS5B interacts with other viral proteins such as core, NS3, 4A, 4B, and 5A. The helicase activity of NS3 seems necessary for RNA strand unwinding during replication, with other nonstructural proteins performing modulatory roles. Cellular proteins interacting with NS5B include VAMP-associated proteins, heIF4AII, hPLIC1, nucleolin, PRK2, ${\alpha}$-actinin, and p68 helicase. The interactions of NS5B with these proteins might play roles in cellular trafficking, signal transduction, and RNA polymerization, as well as the regulation of replication/translation processes.

Concepts concerning various nitrogen efficiencies and their interrelation in rice plant (수도(水稻)에서 여러 질소효율의 개념(槪念)과 상호관계(相互關係))

  • Park, Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.8 no.2
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    • pp.69-80
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    • 1975
  • Relationships between yield and various nitrogen efficiencies, between efficiencies and between efficiency and nitrogen uptake amount of rice plant were proposed and tested using data from N.P.K simple trials about 30 to 50 locations, for three years. Established relationships are well in accordance with experimental results by showing highly significant correlations between them. The overall indications are that high yielding capacity of fields with fertilizer application, depends primarily on high fertilizer nitrogen uptake by increasing fertilizer use efficiency (Eu), secondly the efficiency (Ef) of absorbed fertilizer nitrogen (Nf) and fertilization efficiency (Fe) and also depends much on nitrogen efficiency for grain yield (E) to great extend and that the efficiency (Es) of soil nitrogen (Ns) contributes to E more than Ef does. All nitrogen efficiencies are negatively correlated with the uptake amount of corresponding nitrogen and counterpart efficiency. Es and Ef could be determined firstly by difference method and secondly E versus Cs (Cs=Ns/Ns+Nf) plotting and thirdly E-Cs plotting with labelled fertilizermethod using the equation E=Es Cs+B where B=Ef Cf but a constant under the given condition and at last Y-Ns plotting with labelled fertilizer using Eq Y=$Es{\cdot}Ns+B$ where B=$Ef{\cdot}Nf$. Es which seems not much variable from field to field is mostly greater (about 80% of tested fields) than Ef which is much variable and depends much on fertilizer form. The relationships tested and well agreed are as follows: 1. Y=$Es{\cdot}Ns+Ef{\cdot}Nf$ (Y is yield) 2. E=$Es{\cdot}Cs+Ef{\cdot}Cf$ where Cf=Nf/Nf+Ns 3. E=b-aN where E=E, Es or Ef and N=N, Ns or Nf respectively, (E=Y/N, N=Nf+Ns), b is theoretical maximum under the given system and a is tangent at N=O of the curve, Y=EN. 4. Fe=Ef Eu and Se=$Es{\cdot}Eu$ where Se is efficiency of soil available nitrogen. 5. E=$(Se{\cdot}Cs+Fe{\cdot}Cf)/Eu$ 6. Y=$Es{\cdot}Eu{\cdot}Sf+Ef{\cdot}Eu{\cdot}Fn$or Y=$Es{\cdot}Eu{\cdot}Ea{\cdot}Sn+Ef{\cdot}Eu{\cdot}Fn $where Sf=$Ea{\cdot}Sn$, Ea is soil available nitrogen equivalent to fertilizer(Sf) divided by total soil nitrogen (Sn).

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Numerical Analysis and Simulation for the Pricing of Bond on Term-Structure Interest Rate model with Jump (점프 항을 포함하는 이자율 기간구조 모형의 채권 가격결정을 위한 수치적 분석 및 시뮬레이션)

  • Kisoeb Park
    • Journal of Internet Computing and Services
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    • v.25 no.2
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    • pp.93-99
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    • 2024
  • In this paper, we derive the Partial Differential Bond Price Equation (PDBPE) by using Ito's Lemma to determine the pricing of bond on term-structure of interest rate (TSIR) model with jump. From PDBPE, the Maclaurin series (MS) and the moment-generating function (MGF) for the exponential function are used to obtain a numerical solution (NS) of the bond prices. And an algorithm for determining bond prices using Monte Carlo Simulation (MCS) techniques is proposed, and the pricing of bond is determined through the simulation process. Comparing the results of the implementation of the above two pricing methods, the relative error (RE) is obtained, which means the ratio of NS and MCS. From the results, we can confirm that the RE is less than around 2.2%, which means that the pricing of bond can be predicted very accurately using the proposed algorithms as well as numerical analysis. Moreover, it was confirmed that the bond price obtained using the MS has a relatively smaller error than the pricing of bond obtained by using the MGF.

A Novel IP Forwarding Lookup Scheme for Fast Gigabit IP Routers (초고속 IP 라우터를 위한 새로운 포워딩 Lookup 장치)

  • Kang, Seung-Min;Song, Jae-Won
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.37 no.1
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    • pp.88-97
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    • 2000
  • We have proposed and analysed a novel Lookup Algorithm which had a short switching speed and tiny memory size for IP router. This algorithm could simply be implemeted by a hardware with SRAM because of simple structure. This Lookup scheme needs 1${\sim}$3 memory access times. When we simulated with 40,000 routing record obtained from IPMA Website, the maximum memory size of this algorithm was 316KB(the offset threshold for compression algorithm was 8). When we simulated by HDL using ALTERA EPM7256 series and 100MHz clock and SRAM of 10ns access time, the total lookup time was 45ns for two memory access, 175ns for three memory access.

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On the Properties and Synthesis of Nanostructured W-Cu alloys by Mechanical Alloying(II) Sintering Behavior of MA NS W-Cu Composite Powders (기계적 합금화 방법으로 제조된 nanostructured W-Cu 합금의 제조 및 물성 연구(II) -MA NS W-Cu 복합분말의 소결거동-)

  • 김진천
    • Journal of Powder Materials
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    • v.5 no.2
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    • pp.89-97
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    • 1998
  • Sintering behavior of nanostructured(NS) W-Cu powders prepared by mechanical alloying (MA) was investigated as a function of sintering temperature. MA NS W-2owt%Cu and W-3owt%Cu composite powders with the crystal size of 20-30 nm were annealed at 90$0^{\circ}C$, and thermal characteristics of those powders were investigated by DSC. Sintering behavior of MA NS W-Cu composite powders was investigated during the solid-state sintering and the Cu-liquid phase sintering. The new nanosintering phenonenon of MA W-Cu powders at solid-state sintering temperature was suggested to explain the W-grain growth in the inside of MA powders. The sintering densification of MA NS W-Cu powders was enhanced at Cu melting temperature by arrangement of MA powders, i.e., the first rearrangement of MA powders was occurred, and then the rearrangement of W-grains in the sintered parts was also took place during liquid-phase sintering, i.e., the second rearrangement was happened. Due to the double rearrangement process of MA NS W-Cu powders, the high sintered density with more than 96%o was obtained and the fine and high homogeneous state of W and Cu phases was achieved by sintering at 1200 $^{\circ}C$.

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Characterization of Natural Compounds as Inhibitors of NS1 Endonuclease from Canine Parvovirus Type 2

  • So-Hyung Kwak;Hayeong Kim;Hyeli Yun;Juho Lim;Dong-Hyun Kang;Doman Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.788-796
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    • 2023
  • Canine parvovirus type 2 (CPV-2) has high morbidity and mortality rates in canines. Nonstructural protein 1 (NS1) of CPV-2 has endonuclease activity, initiates viral DNA replication, and is highly conserved. Thus, it is a promising target for antiviral inhibitor development. We overexpressed a 41.9 kDa active recombinant endonuclease in Escherichia coli and designed a nicking assay using carboxyfluorescein and quencher-linked ssDNA as substrates. The optimal temperature and pH of the endonuclease were 37℃ and pH 7, respectively. Curcumin, bisdemethoxycurcumin, demethoxycurcumin, linoleic acid, tannic acid, and α-tocopherol inhibited CPV-2 NS1 endonuclease with IC50 values of 0.29 to 8.03 µM. The extracted turmeric, yerba mate, and sesame cake suppressed CPV-2 NS1 endonuclease with IC50 values of 1.48, 7.09, and 52.67 ㎍/ml, respectively. The binding affinity between curcumin, the strongest inhibitor, and CPV-2 NS1 endonuclease by molecular docking was -6.4 kcal/mol. Curcumin inhibited CPV-2 NS1 endonuclease via numerous hydrophobic interactions and two hydrogen bonds with Lys97 and Pro111 in the allosteric site. These results suggest that adding curcuminoids, linoleic acid, tannic acid, α-tocopherol, extracted turmeric, sesame cake, and yerba to the diet could prevent CPV-2 infection.

An Interferon Resistance Induced by the Interaction between HCV NS5B and Host p48 (C형 간염 바이러스 NS5B 단백질과 숙주의 p48 단백질의 상호작용에 의한 인터페론 저항성의 유도)

  • Park, So-Yeon;Lee, Jong-Ho;Myung, Hee-Joon
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.353-359
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    • 2008
  • Hepatitis C virus (HCV) is known as the causative agent of blood transmitted hepatitis. Two viral proteins, E2 and NS5A, are known to exert interferon resistance of HCV via PKR pathway. Here, we report a third protein, the RNA-dependent RNA polymerase (NS5B) of HCV, induced interferon resistance inhibiting p56 pathway. p56 was shown to interact with p48 subunit of eukaryotic initiation factor 3 (eIF3). This interaction inhibited formation of ternary complex in translation initiation. Using dual reporter assay system, we observed that the translation decreased when interferon alpha was added to the culture. But, in the presence of HCV NS5B, the translation partly recovered. NS5B and p48 subunit of eIF3 were shown to interact. This interaction seems to inhibit the interaction between p48 and p56. This is the first report that a virus exerts interferon resistance via p56 pathway.

Nonsaponin fraction of Korean Red Ginseng attenuates cytokine production via inhibition of TLR4 expression

  • Ahn, Huijeong;Han, Byung-Cheol;Kim, Jeongeun;Kang, Seung Goo;Kim, Pyeung-Hyeun;Jang, Kyoung Hwa;So, Seung Ho;Lee, Seung-Ho;Lee, Geun-Shik
    • Journal of Ginseng Research
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    • v.43 no.2
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    • pp.291-299
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    • 2019
  • Background: Ginsenosides of Korean Red Ginseng extracts (RGE) and its saponin components suppress secretion of inflammasome-mediating cytokines, whereas the nonsaponin fraction (NS) of RGE oppositely stimulates cytokine secretion. Although direct exposure of NS to macrophages in mice induces cytokine production, oral administration of NS has not been studied in inflammasome-related disease in animal models. Methods: Mice were fed RGE or NS for 7 days and then developed peritonitis. Peritoneal cytokines were measured, and peritoneal exudate cells (PECs) were collected to assay expression levels of a set of toll-like receptors (TLRs) and cytokines in response to NS ingestion. In addition, the role of intestinal bacteria in NS-fed mice was assessed. The effect of preexposure to NS in bone marrow-derived macrophages (BMDMs) on cytokine production was further confirmed. Results: NS ingestion attenuated secretion of peritoneal cytokines resulting from peritonitis. In addition, the isolated PECs from NS-fed mice presented lower TLR transcription levels than PECs from control diet-fed mice. BMDMs treated with NS showed downregulation of TLR4 mRNA and protein expression, which was mediated by the $TLR4-MyD88-NF{\kappa}B$ signal pathway. BMDMs pretreated with NS produced less cytokines in response to TLR4 ligands. Conclusion: NS administration directly inhibits TLR4 expression in inflammatory cells such as macrophages, thereby reducing secretion of cytokines during peritonitis.

Analysis of the NS4 Region of Japanese Encephalitis virus K94P05 Isolated from Korea (일본뇌염 바이러스 국내분리주 K94P05의 NS4 부위 분석)

  • Kim, Eun-Jung;Nam, Jae-Hwan;Park, Yong-Kenun;Cho, Hae-Wol
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.197-207
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    • 1997
  • To investigate the NS4 region of JEV, NS4 cDNA of K94P05 (JEV strain isolated from Korea in 1994) was amplified by RT-PCR and analyzed by sequencing PCR product. Genomic size of NS4 was 1212bp and nucleotide sequence was compared with that of other JEV strains. Nucleotide homology between JaOAr582 and K94P05 was 91.1% and that between Beijing and K94P05 was 89.8%, respectively. But the nucleotide sequence of E region of JaOAr582 and K94P05 showed 97.0% homology and that of Beijing and K94P05 did 95.8% homology. NS4 protein was expressed as a form of fusion protein by a prokaryotic expression system. The induced fusion product showed a lower molecular weight than predicted size and remained insoluble. The NS4 protein might be cleavaged by E. coli protease. Concluding above results, high hydrophobicity of the NS4 protein supported the fact that this protein played a role as a membrane component and the poor nucleotide sequence conservativity among JEV strains suggested that this region might be important to adapt each viral growth environment.

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