• Title/Summary/Keyword: MALDI-TOF MS/MS

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Synthesis of Fullerene Oxide by Fullerene ($C_{70}$) and Various Amino N-Oxides under Ultrasonic Irradiation (초음파 조건에서 여러가지 아민 N-옥시드와 풀러렌($C_{70}$)를 이용한 풀러렌 산화물의 합성)

  • Ko, Weon-Bae;Han, Ji-Yeon;Lim, Young-A;Shin, Won-Jic;Yu, Gu-Yong
    • Elastomers and Composites
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    • v.39 no.2
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    • pp.153-160
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    • 2004
  • Fullerene oxides were synthesized by fullerene ($C_{70}$) and several amine N-oxides such as 3-picoline N-oxide, pyridine N-oxide hydrate, quinoline N-oxide, and isoquinoline N-oxide under ultrasonic condition at $25{\sim}43^{\circ}C$. The reactivity of fullerene ($C_{70}$) with various amine N-oxides undo, ultrasonic irradiation showed the same in all of the proceeding experiments; 3-picoline N-oxide : pyridine N-oxide hydrate : quinoline N-oxide : isoquinoline N-oxide. The MALDI-TOF MS, UV-vis spectrophotometer and HPLC analysis confirmed that the products of fullerene oxidation are [$C_{70}(O)n$] (n=1).

The Oxidation of Fullerene[$C_{60}$] using Several Oxidants under Microwave Irradiation (마이크로파 조건에서 여러가지 산화제를 이용한 풀러렌[$C_{60}$의 산화반응)

  • Ko, Weon-Bae;Hwang, Sung-Ho;Ahn, Ju-Hyun
    • Elastomers and Composites
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    • v.40 no.1
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    • pp.45-52
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    • 2005
  • Synthesis or fullerene oxides[$C_{60}(O)_n$] ($n=1{\sim}4$ or n=1) by fullerene[$C_{60}$] and several oxidants such as 3-chloroperoxy benzoic acid, benzoyl peroxide, trichloroisocyanuric acid, and chromium(VI) oxide took place under microwave irradiation. The reactivity in solid state of fullerene[$C_{60}$] with various oxidants under same microwave rendition increased in order or 3- chloroperoxy benzoic acid > benzoyl peroxide > trichloroisocyanuric acid $\simeq$chromium(VI) oxide. The MALDI-TOF-MS, UV-visible spectra and HPLC analysis confirmed that the products of fullerene oxidation were [$C_{60}(O)_n$] ($n=1{\sim}4$ or n=1).

Synthesis of Fullerene Oxides [$C_{70}O_n$] ($n=1{\sim}3$ or n=1) under Microwave Irradiation (마이크로파 조건에서 풀러렌 산화물 [$C_{70}O_n$ ($n=1{\sim}3$ or n=1)의 합성)

  • Ko, Weon-Bae;Ahn, Ju-Hyun;Lim, Young-A;Han, Ji-Yeon;Han, Dong-Sul
    • Elastomers and Composites
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    • v.39 no.4
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    • pp.309-317
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    • 2004
  • Synthesis of fullerene oxides [$C_{70}O_n$] ($n=1{\sim}3$ or n=1) in solid state by fullerene [$C_{70}$] and several oxidants such as 3-chloroperoxy benzoic acid, chromium(VI) oxide, benzoyl peroxide, and trichloroisocyanuric acid was taken place under microwave irradiation. The reactivity in solid state oi fullerene [$C_{70}$] with various oxidants under same microwave condition increased in the order of 3-chloroperoxy benzoic acid > chromium(VI) oxide > trichloroisocyanuric acid ${\simeq}benzoyl$ peroxide. The MALDI-TOF-MS, UV-visible spectra and HPLC analysis confirmed that the products of fullerene oxidation were [$C_{70}O_n$] ($n=1{\sim}3$ or n=1).

The enhancement of protein separation by duplex SDS-PAGE (Duplex SDS-PAGE를 이용한 단백질 분리향상)

  • Pyo, Jae Sung;Roh, Si Hun;Song, Jin-Su;Lee, Kyung Hyeon;Kim, Hie-Joon;Park, Jeong Hill;Kwon, Sung Won
    • Analytical Science and Technology
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    • v.19 no.6
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    • pp.529-534
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    • 2006
  • The protein separation with molecular weight using SDS-PAGE(sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is the one of the most conventional and simple techniques. In, this study, two dimensional SDS-PAGE using same separation principle consecutively was investigated and compared with one dimensional SDS-PAGE. The enhanced separation from duplex SDS-PAGE was observed and separated proteins in the gel were identified by MALDI TOF MS. Identified proteins from different gel spots were found to have different gi numbers. Therefore, duplex SDS-PAGE separation method will be used for economic separation method in the future because only tiny amount of inexpensive reagents are used to perform duplex SDS-PAGE.

Iron Starvation-Induced Proteomic Changes in Anabaena (Nostoc) sp. PCC 7120: Exploring Survival Strategy

  • Narayan, Om Prakash;Kumari, Nidhi;Rai, Lal Chand
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.136-146
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    • 2011
  • This study provides first-hand proteomic data on the survival strategy of Anabaena sp. PCC 7120 when subjected to long-term iron-starvation conditions. 2D-gel electrophoresis followed by MALDI-TOF/MS analysis of iron-deficient Anabaena revealed significant and reproducible alterations in ten proteins, of which six are associated with photosynthesis and respiration, three with the antioxidative defense system, and the last, hypothetical protein all1861, conceivably connected with iron homeostasis. Iron-starved Anabaena registered a reduction in growth, photosynthetic pigments, PSI, PSII, whole-chain electron transport, carbon and nitrogen fixation, and ATP and NADPH content. The kinetics of hypothetical protein all1861 expression, with no change in expression until day 3, maximum expression on the $7^{th}$ day, and a decline in expression from the $15^{th}$ day onward, coupled with in silico analysis, suggested its role in iron sequestration and homeostasis. Interestingly, the up-regulated FBP-aldolase, Mn/Fe-SOD, and all1861 all appear to assist the survival of Anabeana subjected to iron-starvation conditions. Furthermore, the $N_2$-fixation capabilities of the iron-starved Anabaena encourage us to recommend its application as a biofertilizer, particularly in iron-limited paddy soils.

Hydrolytic Degradation of Synthetic Polytrimethylene Terephthalate and Characterization by MALDI-TOF Mass Spectrometry

  • Yang, Eun-Kyung;Jang, Sung-Woo;Cho, Young-Dal;Choe, Eun-Kyung;Park, Chan-Ryang
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.477-482
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    • 2011
  • The structural analysis of polytrimethylene terephthalate (PTT) and characterization of the hydrolytic degradation products after acid hydrolysis were performed using MALDI-TOF mass spectrometry. Mass spectra of the PTT samples were analyzed using a self-calibration method as well as an internal calibration method with standard materials of known masses. PTT structures constituting the samples were determined from the analyses of the spectra, and their relative compositions were estimated. The MALDI-TOF mass spectra of the acid-hydrolyzed PTT sample showed three main series of oligomer products with different end groups in accordance with the hydrolysis schemes. From the spectra of both $Na^+$ and $K^+$ adducts, it was concluded that the PTT samples have higher affinity for $Na^+$ compared with $K^+$ and therefore show higher ionization efficiency with sodium ions when dithranol is used as a matrix. Two different wavelength laser beams ($\lambda$ = 337 nm and 355 nm) were tested and it was observed that the 355 nm beam was more efficient in obtaining the MALDI spectra of PTT using dithranol as a matrix under our experimental conditions.

Identification of Cisplatin-Resistance Associated Genes through Proteomic Analysis of Human Ovarian Cancer Cells and a Cisplatin-resistant Subline

  • Zhou, Jing;Wei, Yue-Hua;Liao, Mei-Yan;Xiong, Yan;Li, Jie-Lan;Cai, Hong-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6435-6439
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    • 2012
  • Chemoresistance to cancer therapy is a major obstacle to the effective treatment of human cancers with cisplatin (DDP), but the mechanisms of cisplatin-resistance are not clear. In this study, we established a cisplatin-resistant human ovarian cancer cell line (COC1/DDP) and identified differentially expressed proteins related to cisplatin resistance. The proteomic expression profiles in COC1 before and after DDP treatment were examined using 2-dimensional electrophoresis technology. Differentially expressed proteins were identified using matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and high performance liquid chromatography-electrospray tandem MS (NanoUPLC-ESI-MS/MS). 5 protein spots, for cytokeratin 9, keratin 1, deoxyuridine triphosphatase (dUTPase), aarF domain containing kinase 4 (ADCK 4) and cofilin1, were identified to be significantly changed in COC1/DDP compared with its parental cells. The expression of these five proteins was further validated by quantitative PCR and Western blotting, confirming the results of proteomic analysis. Further research on these proteins may help to identify novel resistant biomarkers or reveal the mechanism of cisplatin-resistance in human ovarian cancers.

Analysis of Branched PEG-Conjugated Interferon Alpha by Capillary Electrophoresis and MALDI- TOF Mass Spectrometry

  • Na, Dong-Hee;Park, Eun-Ji;Lee, Snag-Deuk;Jo, Young-Woo;Lee, Sung-Hee;Kim, Won-Bae;Lee, Kang-Choon
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.406.3-407
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    • 2002
  • Analysis of interferon alpha (IFN) modified with high molecular weight branched PEG was performed by capillary electrophoresis (CE) and MALDI-TOF mass spectrometry (MALDI-TOF MS). IFN was modified by the reaction of amine residues with an active ester of monomethoxy polyethylene glycol at various molar ratios. As a CE method. capilary electrophoresis sodium dodecyl sulfate nongel sieving (CE-SDS NGS) was performed using an uncoated capilary filled with a hydrophilic replaceable polymer network matrix. (omitted)

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Proteome Analysis of Responses to Ascochlorin in LPS-induced Mouse Macrophage RAW264.7 Cells by 2-D Gel Electrophoresis and MALDI-TOF MS. (LPS로 자극된 macrophage RAW264.7 세포에서 ascochlorin에 대한 단백질체 분석)

  • Chang, Young-Chae
    • Journal of Life Science
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    • v.18 no.6
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    • pp.814-825
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    • 2008
  • Ascochlorin (ASC) is prenyl-phenol compound that was isolated from the fungus Ascochyta viciae. ASC reduces serum cholesterol and triglyceride levels, and suppresses hypertension, tumor development, ameliorates type I and II diabetes. Here, to better understand the mechanisms by which ASC regulates physiological or pathological events and induces responses in the pharmacological treatment of inflammation, we performed differential analysis of the proteome of the mouse macrophage RAW264.7 cells in response to ASC. In this study, we used a proteomic analysis of LPS-induced RAW264.7 cells treated by ASC, to identify proteins potentially involved in inflammatory processes. The RAW264.7 cell proteomes with and without treatment with ASC were compared using two-dimensional electrophoresis (2-D SDS-PAGE), matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF-MS) and bioinformatics. The largest differences in expression were observed for the calreticulin (4-fold decrease), ${\beta}-actin$ (4-fold decrease) and vimentin (1.5-fold decrease). In addition, rabaptin was increased 3-fold in RAW264.7 cells treated with ASC. The expression of some selected proteins was confirmed by RT-PCR analysis.

Toxicoproteomics in the Study of Aromatic Hydrocarbon Toxicity

  • Cho, Chang-Won;Kim, Chan-Wha
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.187-198
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    • 2006
  • The aromatic hydrocarbons (AHs), which include benzene, polycyclic aromatic hydrocarbons, and dioxin, are important chemical and environmental contaminants in industry that usually cause various diseases. Over the years, numerous studies have described and evaluated the adverse health effects induced by AHs. Currently, "Omics" technologies, transcriptomics and proteomics, have been applied in AH toxicity studies. Proteomics has been used to identify molecular mechanisms and biomarkers associated with global chemical toxicity. It could enhance our ability to characterize chemical-induced toxicities and to identify noninvasive biomarkers. The proteomic approach (e.g. 2-dimensional electrophoresis [2-DE]), can be used to observe changes in protein expression during chemical exposure with high sensitivity and specificity. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and electrospray ionization-quadrupole (ESI-Q)-TOF MS/MS are recognized as the most important protein identification tools. This review describes proteomic technologies and their application in the proteomic analysis of AH toxicity.