• Title/Summary/Keyword: Triple resonance

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1H, 15N and 13C Backbone Assignments and Secondary Structures of C-ter100 Domain of Vibrio Extracellular Metalloprotease Derived from Vibrio vulnificus

  • Yun, Ji-Hye;Kim, Hee-Youn;Park, Jung-Eun;Cheong, Hae-Kap;Cheong, Chae-Joon;Lee, Jung-Sup;Lee, Weon-Tae
    • Bulletin of the Korean Chemical Society
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    • v.33 no.10
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    • pp.3248-3252
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    • 2012
  • Vibrio extracellular metalloprotease (vEP), secreted from Vibrio vulnificus, shows various proteolytic function such as prothrombin activation and fibrinolytic activities. Premature form of vEP has an N-terminal (nPP) and a C-terminal (C-ter100) region. The nPP and C-ter100 regions are autocleaved for the matured metalloprotease activity. It has been proposed that two regions play a key role in regulating enzymatic activity of vEP. Especially, C-ter100 has a regulatory function on proteolytic activity of vEP. C-ter100 domain has been cloned into the E. coli expression vectors, pET32a and pGEX 4T-1 with TEV protease cleavage site and purified using gel-filtration chromatography followed by affinity chromatography. To understand how C-ter100 modulates proteolytic activity of vEP, structural studies were performed by heteronuclar multi-dimensional NMR spectroscopy. Backbone $^1H$, $^{15}N$ and $^{13}C$ resonances were assigned by data from standard triple resonance and HCCH-TOCSY experiments. The secondary structures of vEP C-ter100 were determined by TALOS+ and CSI software based on hydrogen/deuterium exchange. NMR data show that C-ter100 of vEP forms a ${\beta}$-barrel structure consisting of eight ${\beta}$-strands.

Fluorescence Resonance Energy Transfer in Calf Thymus DNA from a Long-Lifetime Metal-Ligand Complex to Nile Blue

  • Kang, Jung-Sook;Lakowicz, Josepb R.
    • BMB Reports
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    • v.34 no.6
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    • pp.551-558
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    • 2001
  • We extended the measurable time scale of DNA dynamics to submicrosecond using a long-lifetime metal-ligand complex, $[Ru(phen)_2(dppz)]^{2+}$ (phen=1,10-phenanthroline, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuPD), which displays a mean lifetime near 350 ns. We partially characterized the fluorescence resonance energy transfer (FRET) in calf thymus DNA from RuPD to nile blue (NB) using frequency-domain fluorometry with a high-intensity, blue light-emitting diode (LED) as the modulated light source. There was a significant overlap of the emission spectrum of the donor RuPD with the absorption spectrum of the acceptor NB. The F$\ddot{o}$rster distance ($R_0$) that was calculated from the spectral overlap was $33.4\;{\AA}$. We observed dramatic decreases in the steady-state fluorescence intensities of RuPD when the NB concentration was increased. The intensity decays of RuPD were matched the closest by a triple exponential decay. The mean decay time of RuPD in the absence of the acceptor NB was 350.7 ns. In a concentration-dependent manner, RuPD showed rapid intensity decay times upon adding NB. The mean decay time decreased to 184.6 ns at $100\;{\mu}M$ NB. The FRET efficiency values that are calculated from the mean decay times increased from 0.107 at $20\;{\mu}M$ NB to 0.474 at $100\;{\mu}M$ NB concentration. The use of FRET with a long-lifetime metal-ligand complex donor is expected to offer the opportunity to increase the information about the structure and dynamics of nucleic acids.

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Myocardial viability in acute myocardial infarction: Cardiac MR imaging vs T1 - 201 myocardiac spect

  • 김혜선;박충기;박동우;김용수;김영선;최요원;전석철;서흥석;함창곡
    • Proceedings of the KSMRM Conference
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    • 2002.11a
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    • pp.134-134
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    • 2002
  • 목적: 심근경색의 가역성 여부 판단시 cardiac MR imaging(CMRI)와 myocardial SPECT의 정확성을 비교하기 위하여 대상 및 방법: 흉통을 주소로 내원한 환자 중 CMRI와 SPECT를 시행한 17를 대상으로 역행적으로 조사하였다. CMRI는 double and triple inversion recovery image(IR)에서 심벽의 고신호강도, 2D-FIESTA에서 심기능 분석을 조사하였다. 이어 gadopentetate를 주입하여 확산스캔에서 확산결손을 조사하였다. 연속하여 gadopentetate를 더 주입 후 5분 후 지연심근강조영상을 얻어 지연조영증강 유무와 미세혈류 폐쇄를 조사하였다. SPECT는 아데노신으로 stress 스캔을 얻은 후 T1-201을 재주사 후 rest 스캔하여 확산결손과 재분배를 조사하여 가역성을 판단하였다.

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Wafer-level Vacuum Packaging of a MEMS Resonator using the Three-layer Bonding Technique (3중 접합 공정에 의한 MEMS 공진기의 웨이퍼레벨 진공 패키징)

  • Yang, Chung Mo;Kim, Hee Yeoun;Park, Jong Cheol;Na, Ye Eun;Kim, Tae Hyun;Noh, Kil Son;Sim, Gap Seop;Kim, Ki Hoon
    • Journal of Sensor Science and Technology
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    • v.29 no.5
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    • pp.354-359
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    • 2020
  • The high vacuum hermetic sealing technique ensures excellent performance of MEMS resonators. For the high vacuum hermetic sealing, the customization of anodic bonding equipment was conducted for the glass/Si/glass triple-stack anodic bonding process. Figure 1 presents the schematic of the MEMS resonator with triple-stack high-vacuum anodic bonding. The anodic bonding process for vacuum sealing was performed with the chamber pressure lower than 5 × 10-6 mbar, the piston pressure of 5 kN, and the applied voltage was 1 kV. The process temperature during anodic bonding was 400 ℃. To maintain the vacuum condition of the glass cavity, a getter material, such as a titanium thin film, was deposited. The getter materials was active at the 400 ℃ during the anodic bonding process. To read out the electrical signals from the Si resonator, a vertical feed-through was applied by using through glass via (TGV) which is formed by sandblasting technique of cap glass wafer. The aluminum electrodes was conformally deposited on the via-hole structure of cap glass. The TGV process provides reliable electrical interconnection between Si resonator and aluminum electrodes on the cap glass without leakage or electrical disconnection through the TGV. The fabricated MEMS resonator with proposed vacuum packaging using three-layer anodic bonding process has resonance frequency and quality factor of about 16 kHz and more than 40,000, respectively.

A triple band printed monopole antenna with a bent branch strips for WiFi / 5G (와이파이 및 5G용 굽은 가지 스트립을 가진 삼중대역 인쇄형 모노폴 안테나)

  • Min-Woo Kim;Dong-Gi Shin;Oh-Rim Ryu;Young-Soon Lee
    • Journal of Advanced Navigation Technology
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    • v.25 no.6
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    • pp.536-542
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    • 2021
  • In this paper, we proposed a triple band printed monopole antenna with a bent branch strips for WiFi / 5G. An antenna structure in which bent strips for generating multiple resonance are attached in the form of branches was newly proposed based on a typical monopole strip vertically erected as a triple band antenna structure. The proposed antenna is designed on a FR-4 substrate with dielectric constant 4.3, thickness of 1.6 mm, and size of 28×40 mm2. The measured impedance bandwidth is 430 MHz (2.22~2.65 GHz) in the 2.4 GHz WLAN, 450 MHz (3.38~3.83 GHz) in the 3.5 GHz and 2390 MHz (4.95~7.34 GHz), In particular, it has been observed that antenna has a stable omnidirectional radiation patterns as well as gain of 1.537 dBi, 1.878 dBi and 2.337 dBi in the entire frequency band of interest.

NMR peak assignment for the elucidation of the solution structure of T4 Endonuclease V

  • Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk;Lee, Bong-Jin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.183-183
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    • 1996
  • Bacteriophage T4 endonuclease V initiates the repair of ultraviolet (UV)-induced pyrimidine dimer photoproducts in duplex DNA. The mechanism of DNA strand cleavage involves four sequential stens: linear diffusion along dsDNA, pyrimidine dimer-specific binding,l pyrimidine dimer-DNA glycosylase activity, and Af lyase activity. Although crystal structure is known for this enzyme, solution structure has not been yet known. In order to elucidate the solution structure of this enzyme NMR spectroscopy was used. As a basis for the NMR peak assignment of the protein, HSQC spectrum was obtained on the uniformly $\^$15/N-labeled T4 endonuclease V. Each amide peak of the spectrum were classified according to amino acid spin systems by interpreting the spectrum of $\^$15/N amino acid-specific labeled T4 endonuclease V. The assignment was mainly obtained from three-dimensional NMR spectra such as 3D NOESY-HMQC, 3D TOCSY-HMQC. These experiments were carried out will uniformly $\^$15/N-labeled sample. In order to assign tile resonance of backbon atom, triple-resonance theree-dimensional NMR experiments were also performed using double labeled($\^$15/N$\^$13/C) sample. 3D HNCA, HN(CO)CA, HNCO, HN(CA)HA spectra were recorded for this purpose. The results of assignments were used to interpret the interaction of this enzyme with DNA. HSQC spectrum was obtained for T4 endonuclease V with specific $\^$15/N-labeled amino acids that have been known for important residue in catalysis. By comparing the spectrum of enzyme*DNA complex with that of the enzyme, we could confirm the important role of some residues of Thr, Arg, Tyr in activity. The results of assignments were also used to predict the secondary structure by chemical shift index (CSI).

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Focal nodular hyperplasia: Tripie-contrast enhanced MR imaging using gadolinium chelates, mangafodipir trisodium, and ferumoxides

  • Kim, Joo-Hee;Kim, Myeong-Jin;Park, Young-Nyun;Kim, Kyung-Sik;Lee, Jong-Tae;Yoon, Hyung-Sik
    • Proceedings of the KSMRM Conference
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    • 2001.11a
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    • pp.140-140
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    • 2001
  • We present two cases of surgically proven focal nodular hyperplasia whou underwent tri contrast-enhance MR imaging using gadolinium chelates, mangafodipir trisodium, and ferumoxides After the unehanced MR images were obtained, dynamic gadolinium-enhanced T1-weighted imagi were performed, then mangafodipir enhanced and ferumoxides-enhanced images were obtained. In one case, the mass was isointense on both T1- and T2-weighted images on the unehanced M images, iso and slightly hyperintense on ferumoxides-enhanced FSE and GRE images, strong hyperintense on the mangafodipir enhanced and gadolinium enhanced arterial phase images. In th other case, the mass was isointense on T2-weighted and hypointense on T1-weighted image isointense on ferumoxides-enhanced images, and hyperintense on mangafodipir enhanced an gadolinium enhanced arterial phase images. Triple contrast enhanced MR images were useful correctly diagnose these two cases preoperatively.

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NMR study of the interaction of T4 Endonuclease V with DNA

  • Lee, Bong-Jin;Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.80-80
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    • 1995
  • T4 Endonuclease V (Mw 16,000) acts as a repair enzyme for UV induced pyrimidine dimers in DNA. Many researchers have studied the biochemical characteristics of the enzyme. However the precise action mechanism of T4 endo V has not fully elucidated yet. In our laboratory NMR spectroscopy technique is being used for the structural study of T4 endo V. Because of its low temperature stability and high content of ${\alpha}$-helix, the conventional $^1$H NMR technique was inapplicable. Therefore we utilized stable isotope labeling technique and so far prepared about 10 amino acid specific labeled proteins. The HSQC spectra of amino acid specific labeled proteins will help us to interpret the triple resonance 3D, 4D data which are under processing, We also studied the behaviors of specific amino acid residues whose roles might be critical. When the enzyme labeled by $\^$15/N-Thr was mixed with the substrate oligonucleotide (semispecific -TT- sequence), one crosspeak in its HSQC spectrum was completely desappeared, which means that one of seven Thr residues is in the binding site of the enzyme with DNA, This result is well consistent with previous report that implicated the Thr 2 residue in the activity of the enzyme. Similar studies were carried on the behaviors of Arg and Tyr residues.

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27Al Solid-state NMR Structural Studies of Hydrotalcite Compounds Calcined at Different Temperatures

  • Park, Tae-Joon;Choi, Sung-Sub;Kim, Yong-Ae
    • Bulletin of the Korean Chemical Society
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    • v.30 no.1
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    • pp.149-152
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    • 2009
  • Hydrotalcites are anionic clays that are quite prevalent in nature and their importance is growing more and more because of their very wide range of potential applications and uses. Understanding the structural and compositional changes that occur on the molecular scale during the thermal decomposition of hydrotalcite compounds is essential for the basic prediction and comprehensive understanding of the behavior and technical application of these materials. In this study, several hydrotalcite compounds calcined at different temperatures for applications in a chlorine resistant textile were prepared and 27-Aluminm solid-state nuclear magnetic resonance (NMR) spectroscopy was used as a tool to study their local structure and behavior. The changes in the Al coordination of the hydrotalcite compounds were investigated with one dimensional (1D) solid-state magic angle spinning (MAS) NMR spectroscopy. The two broad resonances arising from the structurally different Al coordinations of these compounds were clearly resolved by two dimensional (2D) triple quantum magic angle spinning (3QMAS) NMR spectroscopy.

Internal Flow and Evaporation Characteristic inside a Water Droplet on a Vertical Vibrating Hydrophobic Surface (수직 진동하는 소수성 표면 위 액적의 내부유동 및 증발특성 연구)

  • Kim, Hun;Lim, Hee-Chang
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.39 no.7
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    • pp.579-589
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    • 2015
  • This study aims to understand the internal flow and the evaporation characteristics of a deionized water droplet subjected to vertical forced vibrations. To predict and evaluate its resonance frequency, the theories of Lamb, Strani, and Sabetta have been applied. To visualize the precise mode, shape, and internal flow inside a droplet, the experiment utilizes a combination of a high-speed camera, macro lens, and continuous laser. As a result, a water droplet on a hydrophobic surface has its typical shape at each mode, and complicated vortices are observed inside the droplet. In particular, large symmetrical flow streams are generated along the vertical axis at each mode, with a large circulating movement from the bottom to the top and then to the triple contact line along the droplet surface. In addition, a bifurcation-shaped flow pattern is formed at modes 2 and 4, whereas a large ellipsoid-shape flow pattern forms at modes 6 and 8. Mode 4 has the fastest internal flow speed and evaporation rate, followed by modes 8 then 6, with 2 having the slowest of these properties. Each mode has the fastest evaporation rate amongst its neighboring frequencies. Finally, the droplet evaporation under vertical vibration would lead to more rapid evaporation, particularly for mode 4.