• Title/Summary/Keyword: In cell NMR

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Expression and Purification of Unstructured Protein, IMUP-1, using Chaperone Co-expression System for NMR Study

  • Yi, Jong-Jae;Yoo, Jung Ki;Kim, Jin Kyeoung;Son, Woo Sung
    • Journal of the Korean Magnetic Resonance Society
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    • v.17 no.1
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    • pp.30-39
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    • 2013
  • Immortalization-upregulated protein-1 (IMUP-1) genes have been cloned and are known to be involved in SV40-mediated immortalization. IMUP-1 gene is highly expressed in various cancer cell lines and tumors, suggesting the possibility that they might be involved in tumorigenicity. Previously, there were several problems for overexpression of IMUP-1 in bacterial expression systems including low solubility and aggregation due to unstructured property. To investigate the structural properties, it is necessary to obtain lots of pure and soluble proteins. Accordingly, the co-expression systems of bacterial chaperone proteins, GroEL-GroES, were used to increase solubility of IMUP-1. From the analysis of NMR and CD experiment data, it is suggested that the protein adopt typical the random coil properties in solution.

Expression and Preparation of Periostin FAS1 Domains for NMR Structure Determination

  • Yun, Hyosuk;Kim, Jae Il;Lee, Chul Won
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.1
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    • pp.17-21
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    • 2016
  • Periostin, a component of extracellular matrix (ECM) protein, is produced and secreted by the fibroblasts that are involved in chronic allergic inflammation diseases and various types of human cancers. Periostin protein is composed of multiple domains including four FAS1 domains which play important roles in cell adhesion and tumor metastasis by interacting with integrins. In spite of their important biological role, the structural information of periosin FAS1 domains was not revealed yet. Recently we systemically prepared various constructs of the FAS1 domains and tried to express them in E. coli. Of them, only single FAS1-II and -IV domains were highly soluble. Circular dichroism (CD) and nuclear magnetic resonance (NMR) studies revealed that the FAS1-IV domain might be suitable for three-dimensional structure determination using NMR spectroscopy.

NMR Analysis of the Substitution of Titanium Tetraisopropoxide with Phenol and Carboxylic Acid

  • Choi, Jeong Chul;Hwang, Kwang-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.21 no.4
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    • pp.135-138
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    • 2017
  • Titanium phenoxide and titanium carboxylate derivatives were prepared by ligand exchange of titanium tetraisopropoxide with the corresponding phenol and carboxylic acids. The substitution reactions were analyzed by NMR focused in the liberation of isopropylalcohol. The chemical shift of secondary proton of isopropyl group shifted to upfield after liberation; from 4.47 ppm at titanium-bound to 4.1~4.3 ppm at free alcohol state in $CDCl_3$. The substitution reaction of titanium tetraisopropoxide with carboxylic acid was applied to form dye-Ti complex for dye-sensitized solar cell.

The Chemical Constituents from Unidentified Sponge (해면의 화학적 성분 연구)

  • Park, Seon Gu;Paul J. Scheuer
    • Journal of the Korean Chemical Society
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    • v.38 no.2
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    • pp.169-173
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    • 1994
  • The previously reported cytotoxic metabolites, against the KB cell line, xestoquinone, halenaquinol sulfate and $halenaquinol^{5,6}$ were isolated from the unidentified sponge collected in October 1992, Manado Bay, Sulawesi in Indonesia. Their structure were elucidated by $^1H-,\;^{13}C$-NMR, $^1H-,\;^{13}C$(1 bond) Heteronuclear Multiple Quantum Coherence Spectroscopy$(HMQC)^1$, $^1H-,\;^{13}C$C(2 and 3 bond) Heteronuclear multiple Bond Correlation Spectroscopy$(HMBC)^2$, Electron Impact Mass Spectroscopy(EI ms), Ultraviolet Spectroscopy(UV), and Infrared Spectroscopy(IR)

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The Chemical Constituents from the Sponge Spongia sp. (해면 Spongia sp.의 화학적 성분 연구)

  • Park, Sun-Ku;Oh, Chang-Sok;Scheuer, Paul-J.
    • Journal of the Korean Chemical Society
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    • v.39 no.4
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    • pp.301-305
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    • 1995
  • The cytotoxic metabolites, against the KB cell line, halenaquinone, epispongiatriol and aldisin were isolated from the sponge Spongia sp. collected in September 1992, Manado Bay, Sulawesi in Indonesia. Their structures were elucidated by 1H, 13C NMR, 1H 13C(1 bond) Heteronuclear Multiple Quantum Coherence Spectroscopy (HMQC), 1H 13C(2 and 3 bond) Heteronuclear Multiple Bond Correlation Spectroscopy (HMBC), Electron Impact Mass Spectroscopy (EI ms) and Infrared Spectroscopy (IR).

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Synthesis and Cytotoxic Activity of (R)-(-)-PGME Amide of Diterpene Acid

  • Lim, Jin-A;Yook, Chan-Nam;You, Il-Soo;Oh, Hyun-Ju;Kim, Myung-Ju;Baek, Seung-Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.520-523
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    • 2005
  • (R)-(-)-PGME amide of diterpene acid (2) was assigned the absolute configuration from NMR correlation experiments. The compound (2) was tested for its growth inhibitory effects against tumor cell lines by the MTT method.

A Cytotoxic Compound from Moutan Cortex Radicis (목단피의 세포독성 물질)

  • Chu, Bo-Yeun;Jin, Wen-Yi;Choi, Su-La;Myung, Pyung-Keun;Seong, Yeon-Hee;Song, Kyung-Sik;Bae, Ki-Hwan
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.3
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    • pp.255-261
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    • 2004
  • The MeOH extracts from 21 species were tested for their cytotoxicity against SK-MEL-28 melanoma cell line and HaCat normal cell line in 5 g/ml by sulforrhodamine-B (SRB) method. Among them, the MeOH extract from Moutan Cortex Radicis showed the moderate activity with the growth rate of 74.3% in SK-MEL-28 cells and the high activity with the growth rate of 207.8% in HaCat cells. Activity-guided fractionation was performed and five compounds, paeonol (1), benzoylpaeoniflorin (2), benzoic acid (3), 2,5-dihydroxy-4-methoxy- acetophenone (4), paeonfliorin (5) were isolated from hexane and EtOAc fraction. The structures were established by physicochemical and spectrometric methods $(mp,\;UV,\;IR,\;^1H-NMR,\;^{13}C-NMR,\;spectram)$. All compounds were evaluated for their cytotoxicity, compound 4 showed the significant cytotoxic activity with $ED_50$ value of $5.92\;{\mu}g/ml$, but the other compounds no activity. These results suggest that compound 4 is a novel anticancer candidate against SK-MEL-28 melanoma cells.

MAS NMR and XRD Study on the Vanadium Site pf Vanadium Silicate Mesoporous Molecular Sieve MCM-41

  • 박동호;Chi-Feng Cheng;Jacek Klinowski
    • Bulletin of the Korean Chemical Society
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    • v.18 no.1
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    • pp.70-75
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    • 1997
  • A wide range (10 < Si/V) of mesoporous vanadium silicate molecular sieves with the MCM-41 structure have been synthesized using vanadyl sulfate as the source of vanadium and characterized by XRD, 51V MAS NMR and 29Si MAS NMR. The increase of the unit cell parameter and the decrease of Q3/Q4 ratio of 29Si spectra with the vanadium content suggest the incorporation of vanadium in the framework of MCM-41 structure. 51V MAS NMR demonstrates that vanadiums in as-synthesized V-MCM-41 are present in the chemical environment of octahedra and octahedral vanadium is decreased and tetrahedral vanadium is increased inversely with raising the calcination temperature. Though the thermal treatment in rotor of hydrated sample resulted in the change from tetrahedral environment to octahedral one and the steaming and the acid treatment affect to the chemical environment of vanadium, the spectrum similar to originally calcined sample is regenerated after recalcination. This indicates that the vanadium is belong to the framework in a relatively exposed site. The best quality XRD pattern of the product of Si/V=27 may be attributable to heterogeneous nucleation mechanism. V-MCM-41's having the Si/V ratio lower than 20 are completely collapsed after calcination.

NMR structural studies on Human CD99 Type I

  • Kim, Hai-Young;Kim, Young-Mee;Joon Shin;Shin, Young-Kee;Park, Seong-Hoe;Lee, Weontae
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.69-69
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    • 2003
  • Human CD99 is a ubiquitous 32-kDa transmembrane protein encoded by the mic2 gene. The major cellular functions of CD99 protein are related to homotypic cell adhension, apoptosis, vesicular protein transport, and differentiation of thymocytes or T cells. Recently it has been reported that expression of a splice variant of CD99 transmembrane protein (Type I and Type II) increases invasive ability of human breast cancer cells. To understand structural basis for cellular functions of CD99 (Type I), we have initiated studies on hCD99$^{TMcytoI}$ and hCD99$^{cytoI}$ using circular dichroism (CD) and multi-dimensional NMR spectroscopy. CD spectrum of hCD99$^{TMcytoI}$ in the presence of 200mM DPC and CHAPS displayed an existence $\alpha$-helical conformation. The solution structure of hCD99$^{cytoI}$ determined by NMR is composed of one N-terminal $\alpha$-helix, $\alpha$A, two C-terminal short $\alpha$-helix segments, $\alpha$B and $\alpha$C. While $\alpha$A and $\alpha$B are connected by the long flexible loop, $\alpha$B and $\alpha$C connected by type III$\beta$-turn. Although it has been rarely figured out the correlation between structure and functional mechanism of hCD99$^{TMcytoI}$ and hCD99$^{cytoI}$, there is possibility of dimerization or oligomerization. In addition, the feasible mechanism of hCD99$^{cytoI}$ is that it could have intramolecular interaction between the N- and C- terminal domain through large flexible AB loop.

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The Cytotoxic Activity of 13(E)-Labd-13-ene-8$\alpha$, 15-diol from Brachyglottis monroi

  • Lim, Jin-A;Kwag, Jung-Sook;Nigel, B.Perry;Baek, Seung-Hwa
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.179.2-179.2
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    • 2003
  • The cytotoxic activity of 13(E)-Labd-13-ene-8$\alpha$, 15-diol(1), isolated from the ethanol extract of Brachyglottis monroi was evaluated against tumor cell lines such as P388, SNU-C4 MDA-MB231, B 16 melanoma and A549 in vitro. By mean of spectral analysis particularly by the aid of various two dimensional NMR experiments, 1H-NMR and 13C-NMR signals of (1) was completely assigned, and thus the structure of (1) was established unambiguously. (omitted)

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