• Title/Summary/Keyword: High-resolution NMR

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Synthesis of Polyrotaxane-biotion Conjugates and Surface Plasmon Resonance Analysis of Streptavidin Recognition

  • Ooya, Tooru;Kawashima, Tomokatsu;Yui, Nobuhiko
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.293-300
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    • 2001
  • A polyrotaxane-biotin conjugate was synthesized and its interaction with streptavidin measured using surface plasmon resonance(SPR) detection. A biodegradable polyrotaxane in which ca, 22 molecules of ${\alpha}$-cyclodextrina(${\alpha}$-CDs) were threaded onto a poly(ethylene oxide) chain(M$\sub$n:4,000) capped with benzyloxycarbonyl-L-phenylalanine was conjugated with a biotin hydorazide and 2-aminoethanol after activing the hydroxyl groups of ${\alpha}$-CDs in the polyrotaxane using N, N'-carbonyldiimidazole. The results of the high-resolution $^1$H-nyclear lmagnetic resonance($^1$H-NMR)spectra and gel permeation chromatography of the conjugate showed that ca, 11 biotin molecules were actually introduced to the polyrotaxane scaffold. An SPR analysis showed that the binding curves of the biotin molecules in the conjugate on the streptavidin-deposited surface changed in a concentration dependent manner, indicating that the biotin in the conjugate was ac-tually recognized by streptavidin. The association equilibrium constant(K$\sub$a/) of the interaction be-tween the conjugate and steptavidin tetramer was of the order 10$\^$7/. These results suggest that polyrotaxane is useful for scaffolds as a polymeric ligand in biomedical fields.

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Isolation and Characterization of Antimicrobial Substance Macrolactin A Produced from Bacillus amyloliquefaciens CHO104 Isolated from Soil

  • Lee, Seung-Je;Cho, Jeong-Yong;Cho, Jung-Il;Moon, Jae-Hak;Park, Ki-Deok;Lee, Young-Ju;Park, Keun-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.525-531
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    • 2004
  • A strain antagonistic to Fusarium solani, CHO104, was selected from approximately 100 microorganisms isolated from soil. Strain CHO104 was identified as Bacillus amyloliquefaciens and found to be Gram-positive based on the Biolog system and 16S rRNA methods. A culture broth of B. amyloliquefaciens CHO104 also exhibited antimicrobial activity against various microorganisms. As such, the EtOAc extract of the culture broth was isolated by various column chromatographic procedures and HPLC. The antimicrobial and antifungal substance was then characterized as macrolactin A $(C_{24}H_{34}O_5)$ using high-resolution EI-MS and NMR analyses, and found to be very effective in inhibiting the growth of Staphylococcus aureus, E. coli, and Botrytis cinerea, even when using a concentration of one-twentieth of the benzoic acid as the control compound.

Rescuing p53 from mdm2 by a pre-structured motif in intrinsically unfolded SUMO specific protease 4

  • Kim, Do-Hyoung;Lee, Chewook;Kim, Bom;Lee, Si-Hyung;Han, Kyou-Hoon
    • BMB Reports
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    • v.50 no.10
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    • pp.485-486
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    • 2017
  • Many intrinsically unstructured/unfolded proteins (IUPs) contain transient local secondary structures even though they are "unstructured" in a tertiary sense. These local secondary structures are named "pre-structured motifs (PreSMos)" and in fact are the specificity determinants for IUP-target binding, i.e., the active sites in IUPs. Using high-resolution NMR we have delineated a PreSMo active site in the intrinsically unfolded mid-domain (residues 201-300) of SUMO-specific protease 4 (SUSP4). This 29-residue motif which we termed a p53 rescue motif can protect p53 from mdm2 quenching by binding to the p53-helix binding pocket in mdm2(3-109). Our work demonstrates that the PreSMo approach is quite effective in providing a structural rationale for interactions of p53-mdm2-SUSP4 and opens a novel avenue for designing mdm2-inhibiting anticancer compounds.

A Tumor Growth Inhibitory Substance Isolated from Panax ginseng (고려홍삼분말중의 항종양 활성물질)

  • Katano Mitsuo;Yamamoto Hiroshi;Matsunaga Hisashi
    • Proceedings of the Ginseng society Conference
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    • 1988.08a
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    • pp.33-35
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    • 1988
  • During a series of studies aimed at isolation of the tumor growth inhibitory substance from Panax ginseng. we found a new type of antitumor substance. The substance was isolated from a powder of the root of Panax ginseng C.A. Meyer. which is commonly used for various diseases as a commercial medical drug by the name of Korean Red Ginseng Powder in Japan. Data from infrared spectra proton and carbon-13 nuclear magnetic resonance. and high resolution mass spectra were identical with those of panaxytriol. Panaxytriol isolated from Korean Red Ginseng Powder (Nikkan Korai Ninjin Co.. Ltd.. Japan) inhibited the growth of several kinds of human and murine malignant cells in vitro. Although the detailed mechanism of cell growth inhibition by panaxytriol is yet to he elucidated. panaxytriol's action appeares to be more dose-dependent than time-dependent.

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RK-270D and E, Oxindole Derivatives from Streptomyces sp. with Anti-Angiogenic Activity

  • Jang, Jun-Pil;Jang, Mina;Nogawa, Toshihiko;Takahashi, Shunji;Osada, Hiroyuki;Ahn, Jong Seog;Ko, Sung-Kyun;Jang, Jae-Hyuk
    • Journal of Microbiology and Biotechnology
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    • v.32 no.3
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    • pp.302-306
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    • 2022
  • A chemical investigation of a culture extract from Streptomyces sp. RK85-270 led to the isolation and characterization of two new oxindoles, RK-270D (1) and E (2). The structures of 1 and 2 were determined by analyzing spectroscopic and spectrometric data from 1D and 2D NMR and High-resolution electrospray ionization mass spectrometry (HRESIMS) experiments. Compound 1 exhibited anti-angiogenic activities against human umbilical vein endothelial cells (HUVECs) without cytotoxicity. Results of Western blot analysis revealed that 1 inhibits VEGF-induced angiogenesis in the HUVECs via VEGFR2/ p38 MAPK-mediated pathway.

Non-monotonic Size Dependence of Electron Mobility in Indium Oxide Nanocrystals Thin Film Transistor

  • Pham, Hien Thu;Jeong, Hyun-Dam
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2505-2511
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    • 2014
  • Indium oxide nanocrystals ($In_2O_3$ NCs) with sizes of 5.5 nm-10 nm were synthesized by hot injection of the mixture precursors, indium acetate and oleic acid, into alcohol solution (1-octadecanol and 1-octadecence mixture). Field emission transmission electron microscopy (FE-TEM), High resolution X-Ray diffraction (X-ray), Nuclear magnetic resonance (NMR), and Fourier transform infrared spectroscopy (FT-IR) were employed to investigate the size, surface molecular structure, and crystallinity of the synthesized $In_2O_3$ NCs. When covered by oleic acid as a capping group, the $In_2O_3$ NCs had a high crystallinity with a cubic structure, demonstrating a narrow size distribution. A high mobility of $2.51cm^2/V{\cdot}s$ and an on/off current ratio of about $1.0{\times}10^3$ were observed with an $In_2O_3$ NCs thin film transistor (TFT) device, where the channel layer of $In_2O_3$ NCs thin films were formed by a solution process of spin coating, cured at a relatively low temperature, $350^{\circ}C$. A size-dependent, non-monotonic trend on electron mobility was distinctly observed: the electron mobility increased from $0.43cm^2/V{\cdot}s$ for NCs with a 5.5 nm diameter to $2.51cm^2/V{\cdot}s$ for NCs with a diameter of 7.1 nm, and then decreased for NCs larger than 7.1 nm. This phenomenon is clearly explained by the combination of a smaller number of hops, a decrease in charging energy, and a decrease in electronic coupling with the increasing NC size, where the crossover diameter is estimated to be 7.1 nm. The decrease in electronic coupling proved to be the decisive factor giving rise to the decrease in the mobility associated with increasing size in the larger NCs above the crossover diameter.

Measurement and Assessment of Absolute Quantification from in Vitro Canine Brain Metabolites Using 500 MHz Proton Nuclear Magnetic Resonance Spectroscopy: Preliminary Results (개의 뇌 조직로부터 추출한 대사물질의 절대농도 측정 및 평가: 500 MHz 고자장 핵자기공명분광법을 이용한 예비연구결과)

  • Woo, Dong-Cheol;Bang, Eun-Jung;Choi, Chi-Bong;Lee, Sung-Ho;Kim, Sang-Soo;Rhim, Hyang-Shuk;Kim, Hwi-Yool;Choe, Bo-Young
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.2
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    • pp.100-106
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    • 2008
  • The purpose of this study was to confirm the exactitude of in vitro nuclear magnetic resonance spectroscopy(NMRS) and to complement the defect of in vivo NMRS. It has been difficult to understand the metabolism of a cerebellum using in vivo NMRS owing to the generated inhomogeneity of magnetic fields (B0 and B1 field) by the complexity of the cerebellum structure. Thus, this study tried to more exactly analyze the metabolism of a canine cerebellum using the cell extraction and high resolution NMRS. In order to conduct the absolute metabolic quantification in a canine cerebellum, the spectrum of our phantom included in various brain metabolites (i.e., NAA, Cr, Cho, Ins, Lac, GABA, Glu, Gln, Tau and Ala) was obtained. The canine cerebellum tissue was extracted using the methanol-chloroform water extraction (M/C extraction) and one group was filtered and the other group was not under extract processing. Finally, NMRS of a phantom solution and two extract solution (90% D2O) was progressed using a 500MHz (11.4 T) NMR machine. Filtering a solution of the tissue extract increased the signal to noise ratio (SNR). The metabolic concentrations of a canine cerebellum were more close to rat’s metabolic concentration than human’s metabolic concentration. The present study demonstrates the absolute quantification technique in vitro high resolution NMRS with tissue extraction as the method to accurately measure metabolite concentration.

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Ex Vivo ${1}^H$ MR Spectroscopy: Normal gastric and cancer tissue (정상 위 조직과 위암 조직의 시험관 내 수소자기공명분광)

  • Cho Ji Youn;Shin Oon Jae;Choi Ki Seung;Kim Su Hyun;Eun Choong Ki;Yang Young Il;Lee Jung Hee;Mun Chi Woong
    • Journal of Gastric Cancer
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    • v.3 no.3
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    • pp.151-157
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    • 2003
  • Purpose: In this study, we attempted to ascertain the proton magnetic resonance spectroscopy (${1}^H$ MRS) peak characteristics of human gastric tissue layers and finally to use the metabolic peaks of MRS to distinguish between normal and abnormal gastric specimens. Materials and Methods: Ex-vivo ${1}^H$ MRS examinations of thirty-five gastric specimens were performed to distinguish abnormal gastric tissues invaded by carcinoma cells from normal stomach-wall tissues. High-resolution 400-MHz (9.4-T) ${1}^H$ nuclear magnetic resonance (NMR) spectra of two gastric layers, a proper muscle layer, and a composite mucosasubmucosa layer were compared with those of clinical 64- MHz (1.5-T) MR spectra. Three-dimensional spoiled gradient recalled (SPGR) images were used to determine the size and the position of a voxel for MRS data collection. Results: For normal gastric tissue layers, the metabolite peaks of 400-MHz ${1}^H$ MRS were primarily found to be as follows: lipids at 0.9 ppm and 1.3 ppm; alanine at 1.58 ppm; N-acetyl neuraminic acid (sialic acid) at 2.03 ppm; and glutathione at 2.25 ppm in common. The broad and featureless featureless spectral peaks of the 64-MHz MRS were bunched near 0.9, 1.3, and 2.0, and 2.2 ppm in human specimens without respect to layers. In a specimen (Borrmmann type III) with a tubular adenocarcinoma, the resonance peaks were measured at 1.26, 1.36 and 3.22 ppm. All the peak intensities of the spectrum of the normal gastric tissue were reduced, but for gastric tumor tissue layers, the lactate peak split into 1.26 and 1.39 ppm, and the peak intensity of choline at 3.21 ppm was increased. Conclusion: We found that decreasing lipids, an increasing lactate peak that split into two peaks, 1.26 ppm and 1.36 ppm, and an increasing choline peak at 3.22 ppm were markers of tumor invasion into the gastric tissue layers. This study implies that MR spectroscopy can be a useful diagnostic tool for gastric cancer.

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Quantitation of L-carnitine in plasma by electrospray ionization tandem mass spectrometry (ESI/MS/MS를 이용한 혈장 중 카르니틴 정량분석)

  • Kang, Seung Woo;Kim, Ho Hyun;Lee, Kyung Ryul;Yoon, Hye-Ran
    • Analytical Science and Technology
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    • v.18 no.2
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    • pp.163-167
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    • 2005
  • In this study, a novel analytical method has been developed for the rapid determination of L-carnitine in human plasma using electrospray ionization tandem mass spectrometry. Free carnitine (FC) was analyzed after extraction with 80% methanol and total carnitine (TC) was analyzed after hydrolysis and extraction. Acyl carnitine (AC) was subtracted FC from TC. Analytical methods used multiple reaction monitoring (MRM) scan modes. A correlation coefficient of linear regression ($r^2$) was 0.9995, recovery was 97%, reproducibility was less than 10%, and limit of detection (LOD) was $0.0016{\mu}mol/L$. This method reduced sample preparation time and showed high resolution and good reproducibility compared to that with liquid chromatographic methods. Normal control showed AC was lower than FC. Clinical management of patients with inborn error of metabolism showed FC was lower than AC. Thus, carnitine fraction level was very important to monitoring patients with metabolic disorder.

Synthesis and Characterization of Bis-Thienyl-9,10-anthracenes Containing Electron Withdrawing 2-Cyanoacrylic Acid or 2-Methylenemalononitrile Group

  • Wang, Yuan;Yu, Qu Feng;Park, Hea-Jung;Ryu, Suk-Hwa;Choi, Jung-Hei;Yoon, Ung-Chan
    • Bulletin of the Korean Chemical Society
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    • v.32 no.spc8
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    • pp.3081-3089
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    • 2011
  • A series of new bis-thienylanthracene derivatives D1~D5 containing 9,10-antharcene moiety in the center and 2-methylenemalonotitrile or 2-cyanoacrylic acid functional group on the terminal thiophenes were synthesized and characterized by $^1H$-NMR and high-resolution mass spectroscopy. Their optical, electrochemical, and thermal properties were measured. They have absorption ${\lambda}_{max}$ in the range of 437~480 nm and max of $7.4{\times}10^3{\sim}2.0{\times}10^4M^{-1}cm^{-1}$. The substitution of 2-cyanoacrylic acid group allows greater value of ${\varepsilon}_{max}$ than that of 2-methylenemalonotitrile. TGA curves showed that D4 and D5 which have 2-cyanoacrylic acid functional group on the terminal thiophene(s) exhibit good thermal stability and D4 was thermally stable up to $400^{\circ}C$. Their optical properties and LUMO energy levels measured suggest that they can serve as potential candidates for electron donor materials of organic photovoltaic cells (OPVs) or D4 and D5 which contain 2-cyanoacrylic acid group can be used as organic dyes of dye-sensitized solar cells (DSSCs).