• Molecules and Cells
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• 제22권1호
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• pp.13-20
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• 2006

Hepatitis C virus (HCV) non-structural protein 5A protein (NS5A), which consists of three functional domains, is involved in regulating viral replication, interferon resistance, and apoptosis. Recently, the three-dimensional structure of the domain 1 was determined. However, currently the molecular basis for the domains 2 and 3 of HCV NS5A is yet to be defined. Toward this end, we expressed, purified the domain 2 of the NS5A (NS5A-D2), and then performed biochemical and structural studies. The purified domain 2 was active and was able to bind NS5B and PKR, biological partners of NS5A. The results from gel filtration, CD analysis, 1D $^1H$ NMR and 2D $^1H-^{15}N$ heteronuclear single quantum correlation (HSQC) spectroscopy indicate that the domain 2 of NS5A appears to be flexible and disordered.

• BMB Reports
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• 제37권6호
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• pp.741-748
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• 2004

The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.

• Journal of Microbiology and Biotechnology
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• 제29권10호
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• pp.1665-1674
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• 2019

Zika virus (ZIKV) is a mosquito-transmitted, emerging Flavivirus that causes Guillain-$Barr{\acute{e}}$ syndrome and microcephaly in adults and fetuses, respectively. Since ZIKV was first isolated in 1947, severe outbreaks have occurred at various places worldwide, including Yap Island in 2007, French Polynesia in 2013, and Brazil in 2015. Although incidences of ZIKV infection and dissemination have drastically increased, the mechanisms underlying the pathogenesis of ZIKV have not been sufficiently studied. In addition, despite extensive research, the exact roles of individual ZIKV genes in the viral evasion of the host innate immune responses remain elusive. Besides, it is still possible that more than one ZIKV-encoded protein may negatively affect type I interferon (IFN) induction. Hence, in this study, we aimed to determine the modulations of the IFN promoter activity, induced by the MDA5/RIG-I signaling pathway, by over-expressing individual ZIKV genes. Our results show that two nonstructural proteins, NS2A and NS4A, significantly down-regulated the promoter activity of IFN-${\beta}$ by inhibiting multiple signaling molecules involved in the activation of IFN-${\beta}$. Interestingly, while NS2A suppressed both full-length and constitutively active RIG-I, NS4A had inhibitory activity only on full-length RIG-I. In addition, while NS2A inhibited all forms of IRF3 (full-length, regulatory domain-deficient, and constitutively active), NS4A could not inhibit constitutively active IRF3-5D. Taken together, our results showed that NS2A and NS4A play major roles as antagonists of MDA5/RIG-I-mediated IFN-${\beta}$ induction and more importantly, these two viral proteins seem to inhibit induction of the type I IFN responses in differential mechanisms. We believe this study expands our understanding regarding the mechanisms via which ZIKV controls the innate immune responses in cells and may pave the way to development of ZIKV-specific therapeutics.

• 한국광학회지
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• 제19권5호
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• pp.370-375
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• 2008

본 논문에서 비동시성 광샘플링(asynchronous optical sampling; AOS) 방식을 이용하는 고속 고분해 테라헤르츠 시간영역 분광(terahertz time domain spectroscopy; THz-TDS)을 시연한다. 모터로 구동되는 선형 스테이지를 사용하지 않고, 약간 다른 반복 주파수를 갖는 두 대의 펨토초 레이저를 각각 테라헤르츠파 발생과 검출에 사용하여 고속으로 10 ns의 시간축 상의 신호를 획득하고 fast Fourier transformation(FFT)을 통하여 100 MHz의 주파수 분해능을 갖는 고분해 분광을 구현한다. Cross-correlation 방법에 의해 시간 분해능은 278 fs으로 측정되었다. 또한, 본 분광기를 이용하여 수증기의 투과 스펙트럼을 측정하고 흡수선들을 분석하였다.

• 한국광학회지
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• 제23권5호
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• pp.211-216
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• 2012

본 논문은 분광 영역 모드록킹(Fourier domain mode-locking: FDML) 레이저를 기반으로 공진형 광섬유 격자 센서를 구현한 결과를 보고한다. FDML 레이저는 파브리-페롯 가변 필터를 이용하여 링 형태로 구성하며, 레이저 공진기 안에 광섬유 격자 2개를 한 쌍으로 하여 센서부 2개를 삽입한 구조이다. 광섬유 격자는 반사 거울 역할을 하며, 광섬유 격자의 위치에 따라 독립된 FDML 레이저 공진기를 구성한다. 각각 센서부의 공진 주파수는 46.687 kHz 와 44.340 kHz이다. FBG 센서 시스템에 정적 및 동적 스트레인을 가하였으며, 정적 스트레인에 대해 파장 영역과 시간 영역에서 측정된 스트레인에 대한 변화율은 각각 $0.61pm/{\mu}{\epsilon}$, $0.8ns/{\mu}{\epsilon}$ 이다.

• Genomics & Informatics
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• 제19권4호
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• pp.48.1-48.10
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• 2021

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) encodes small envelope protein (E) that plays a major role in viral assembly, release, pathogenesis, and host inflammation. Previous studies demonstrated that pyrazine ring containing amiloride analogs inhibit this protein in different types of coronavirus including SARS-CoV-1 small envelope protein E (SARS-CoV-1 E). SARS-CoV-1 E has 93.42% sequence identity with SARS-CoV-2 E and shared a conserved domain NS3/small envelope protein (NS3_envE). Amiloride analog hexamethylene amiloride (HMA) can inhibit SARS-CoV-1 E. Therefore, we performed molecular docking and dynamics simulations to explore whether amiloride analogs are effective in inhibiting SARS-CoV-2 E. To do so, SARS-CoV-1 E and SARS-CoV-2 E proteins were taken as receptors while HMA and 3-amino-5-(azepan-1-yl)-N-(diaminomethylidene)-6-pyrimidin-5-ylpyrazine-2-carboxamide (3A5NP2C) were selected as ligands. Molecular docking simulation showed higher binding affinity scores of HMA and 3A5NP2C for SARS-CoV-2 E than SARS-CoV-1 E. Moreover, HMA and 3A5NP2C engaged more amino acids in SARS-CoV-2 E. Molecular dynamics simulation for 1 ㎲ (1,000 ns) revealed that these ligands could alter the native structure of the proteins and their flexibility. Our study suggests that suitable amiloride analogs might yield a prospective drug against coronavirus disease 2019.

• Journal of Photoscience
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• 제11권3호
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• pp.133-139
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• 2004

The metal-ligand complex, $[Ru(phen)_2(dppz)]^{2+}$ (phen=1,10-phenanthroline, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuPD), was used as a spectroscopic probe for studying nucleic acid dynamics. The RuPD complex displays a long lifetime and a molecular light switch property upon DNA binding due to shielding of its dppz ligand from water. To show the usefulness of this luminophore (RuPD) for probing nucleic acid dynamics, we compared its intensity and anisotropy decays when intercalated into the $tRNA^{val}$ and pBluescript (pBS) II SK(+) phagemid through a comparison with ethidium bromide (EB), a conventional nucleic acid probe. We used frequency-domain fluorometry with a blue light-emitting diode (LED) as the modulated light source. The mean lifetime for the $tRNA^{val}$ (<${\tau}$> = 166.5 ns) was much shorter than that for the pBS II SK(+) phagemid (<${\tau}$> = 481.3 ns), suggesting a much more efficient shielding from water by the phagemid. Because of their size difference, the anisotropy decay data showed a much shorter rotational correlation times for the $tRNA^{val}$ (99.9 and 23.6 ns) than for the pBS II SK(+) phagemid (968.7 and 39.5 ns). These results indicate that RuPD can be useful for studying nucleic acid dynamics.

• Archives of Pharmacal Research
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• 제25권2호
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• pp.143-150
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• 2002

Fluorescent probes bound to DNA typically display nanosecond decay times and reveal only nanosecond motions. We extend the time range of measurable DNA dynamics using $[Ru(pby)_2(dppz)]^{2+}$ (bpy=2.2'-bipyridine, dppz=dipyrido[3,2-a2',3'-c]phenazine) (RuBD) which displays a mean lifetime near 90 ns. To test the usefulness of RuBD as a probe for diffusive processes in calf thymus DNA, we compared the efficiencies of fluorescence resonance energy transfer (FRET) using three donors which display lifetimes near 5 ns for acridine orange (AO), 22 ns for ethidum bromide (EB) and 92 ns for RuBD, with nile blue (NB) as the acceptor. The F rster distances for AO-NB, EB-NB and RuBD-NB donor-acceptor pairs were 42.3, 52.3, and $30.6{\;}{\AA}$, respectively. All three donors showed dramatic decreases in fluorescence intensities and more rapid intensity decays with increasing NB concentrations. The intensity decays of AO and EB in the presence of varying concentrations of NB were satisfactorily described by the one-dimensional FRET model without diffusion (Blumen and Manz, 1979). In the case of the long-lifetime donor RuBD, the experimental phase and modulation somewhat deviated from the recovered values computed from this model. The recovered NB concentrations and FRET efficiencies from the model were slightly larger than the expected values, however, the recovered and expected values did not show a significant difference. Thus, it is suggested that the lifetime of RuBD is too short to measure diffusive processes in calf thymus DNA.

• 한국자기학회지
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• 제17권5호
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• pp.188-193
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• 2007

[ $1.00{\times}0.24\;{\mu}m^2$ ] 크기의 $Ni_{80}Fe_{20}$ 박막의 자화 반전 거동을 자화 용이축으로 1 ns 이하의 펄스 자기장의 지속 시간과 세기를 변수로 인가하여 micromagnetics 시뮬레이션으로 관찰하였다. 자성 박막은 직사각형과 타원형의 모양을 가지며, 두께는 2 nm와 4 nm로 설정하였다. 실험 결과 $Ni_{80}Fe_{20}$ 박막의 두께와 모양에 따라 각각 다른 경향을 보이는 것을 확인할 수 있었다. 박막의 두께에 따라 두께 방향으로 형성되는 반자장의 크기 차에 의해 edge domain에서 스핀의 회전속도와 스핀 스위칭의 거동에 차이가 생기며, 박막이 두꺼울수록 자화 반전에 더 긴 펄스 지속 시간과 강한 펄스 자기장이 필요하다는 것을 확인하였다. 한편, 자화 반전이 예상되는 영역에서 자화 반전이 일어나지 않는 비정상적인 자화 반전 영역을 발견할 수 있었는데, 박막의 모양이 타원일 때와 박막의 두께가 얇은 경우에 그 영역이 더욱 불규칙적이고, 넓게 분포하였다. 이러한 현상은 막의 두께가 매우 얇기 때문에 두께 방향으로 형성된 강한 반자장의 영향에 의해 나타나는 것으로 여겨진다. Edge domain이 더 많은 직사각형 모양의 경우 자화반전이 일어나는 동안 자기모멘트의 세차 운동에 의해 생기는 $M_z$ 성분, 즉 두께방향으로 형성된 반자장이 더 작고, 이에 따라 자화 반전이 예상되는 영역에서 자화 반전이 일어나지 않는 비정상적인 자화 반전 영역이 더 적어짐을 확인하였다. 본 시뮬레이션 결과는 자성박막의 안정된 고속 자화반전을 위해서는 반자장의 영향을 최소화하는 것이 중요하다는 것을 보여준다.

• 비파괴검사학회지
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• 제24권5호
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• pp.499-507
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• 2004

광섬유 ROTDR (Rayleigh optical time domain reflectometry) 센서와 보디 긴 광섬유를 감지광섬유로 사용할 수 있는 광섬유 BOTDA (Brillouin optical time domain analysis) 센서를 구성하고, 이들 각각을 이용하여 중요보안 대상체인 사회기반시설물에 침투하는 침입자를 탐지할 수 있는 기포 연구를 수행하였다 ROTDR 센서의 감지부로는 넓은 면적을 감지할 수 있는 매설형 광섬유 센서 탐지판을 제작하고, 인가된 침입물체의 위치와 무게에 따른 신호특성을 고찰하였다. ROTDR 센서는 펄스 폭이 30ns이고, 광섬유의 길이는 10km 이상이다. 위치탐지오차는 약 2m 이내였으며, 무게에 따른 탐지능력은 20kgf, 40kgf, 60kgf, 80 kgf의 네 단계를 구분할 수 있음을 알 수 있었다. 넓은 지역에 걸친 침입자의 침투를 감시하기 위하며 수십 km의 광섬유 길이 전제를 감지부로 사용할 수 있는 광섬유 BOTDA 센서를 개발하였다. BOTDA 센서는 한 개의 레이저 다이오드와 두 개의 광전 변조기(electro-optic modulator)를 사용하여 간단하게 구성하였다. 침입자에 의한 광섬유의 변형률 벽화를 탐지하는 실험을 수행하기 위하여 광학테이블 위에 광섬유에 변형률을 인가하기 위한 실험 장치를 설치하여 실험을 수행하였다. 이 실험으로부터 시간간격 1.5 초동안 광섬유 약 4.81km의 길이를 거리분해능 3m로 침입자를 탐지할 수 있음을 확인하였다.