• Title/Summary/Keyword: ion trap mass spectrometry

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Proteomics Analysis of Gastric Epithelial AGS Cells Infected with Epstein-Barr Virus

  • Ding, Yong;Li, Xiao-Rong;Yang, Kai-Yan;Huang, Li-Hua;Hu, Gui;Gao, Kai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.367-372
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    • 2013
  • Effects of the Epstein-Barr virus (EBV) on cellular protein expression are essential for viral pathogenesis. To characterize the cellular response to EBV infection, differential proteomes of gastric epithelial AGS cells were analyzed with two-dimensional gel electrophoresis (2-DE) followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and liquid chromatography electrospray/ionization ion trap (LC-ESI-IT) mass spectrometry identification. Mass spectrometry identified 9 altered cellular proteins, including 5 up-regulated and 4 down-regulated proteins after EBV infection. Notably 2-DE analysis revealed that EBV infection induced increased expression of heat shock cognate 71 kDa protein, actin cytoplasmic 1, pyridoxine-5'-phosphate oxidase, caspase 9, and t-complex protein 1 subunit alpha. In addition, EBV infection considerably suppressed those cellular proteins of zinc finger protein 2, cyclin-dependent kinase 2, macrophage-capping protein, and growth/differentiation factor 11. Furthermore, the differential expressional levels of partial proteins (cyclin-dependent kinase 2 and caspase 9) were confirmed by Western blot analysis.Thus, this work effectively provided useful protein-related information to facilitate further investigation of the mechanisms underlying EBV infection and pathogenesis.

Tandem Mass Spectrometry of N-linked Glycans from Human Immunoglobulin G (다중 질량 분석법을 이용한 인체 면역글로불린 G의 N-연결 글라이칸 분석)

  • Joo, Hwang-Soo;Kim, Yun-Gon;Jang, Kyoung-Soon;Kim, Byung-Gee
    • KSBB Journal
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    • v.22 no.4
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    • pp.234-238
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    • 2007
  • We used electrospary ionization ion trap tandem mass spectrometry (ESI-IT tandem MS) to structural elucidation of three different biantennary-type glycans having zero, one, two galactoses (G0, G1, G2). The highest fragment ion in the MS/MS spectra of three glycans was produced by 0,2-ring cleavage of fucose-linked N-acetylglucosamine (GlcNAc) in reducing end. The fragment ions both from precursor ions and 0,2-ring cleaved ions ($^{0.2}An$; n=5 for G0, n=6 for G1 and G2) were not overlapped each other. As results of $MS^n$ analyses, tandem fragmentation trees of each glycans were generated and 2,4-ring cleavages ($^{2.4}A_6$) were occurred in GlcNAc linked to reducing end GlcNAc. This structural elucidation and fragmentation study of N-linked glycans by tandem mass spectrometry can be applied to structural analysis of more complicated glycans.

Analytical Method of Epichlorohydrin in Canned Beverages by Purge-and- Trap/GC

  • Lee Kwang-Ho;Kwak In-Shin;Kim Dyoung-Il;Choi Byoung-Hee;Kim Guy-Joung;Lee Chul-Won
    • Proceedings of the Korean Society of Food Hygiene and Safety Conference
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    • 2001.10a
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    • pp.140-140
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    • 2001
  • A sensitive analytical method based on gas chromatograpy-mass spectrometry with a selected ion monitoring (GC/MS-SIM) with the purge-and-trap concentration and with headspace method (in limited applications) was developed for determining of epichlorohydrin in canned beverages coated with epoxy resin. The calibration curve in the range of $0.5\sim50ng$ had correlation coefficient greater than 0.998 and a detection limit of $0.l\mug/L$ was obtained using a sample volume of 20ml. The predominant ions of epichlorohydrin produced in MSD using electron ionization(EI) were m/z 57 ([M-CI]+) and 62/64 $([M-CH_2O]+)$. In survey of epichlorohydrin in thirty commercial canned beverage samples, none of them was detected.

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Comparative Proteome Analysis of Cyanidin 3-O-glucoside Treated Helicobacter pylori

  • Kim, Sa-Hyun;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.21 no.4
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    • pp.233-240
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    • 2015
  • Some virulence proteins of Helicobacter pylori, such as vacuolating cytotoxic protein A (VacA) and cytotoxin-associated gene protein A (CagA) have been reported to be causative agents of various gastric diseases including chronic gastritis, gastric ulcer or gastric adenocarcinoma. The expression level of these virulence proteins can be regulated when H. pylori is exposed to the antibacterial agent, cyanidin 3-O-glucoside (C3G) as previously reported. In this study, we analyzed the quantitative change of various virulence proteins including CagA and VacA by C3G treatment. We used 2-dimensional electrophoresis (2-DE) to analyze the quantitative change of representative ten proteome components of H. pylori 60190 ($VacA^+/CagA^+$; standard strain of Eastern type). After 2-DE analysis, spot intensities were analyzed using ImageMaster$^{TM}$ 2-DE Platinum software then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). Next, we selected major virulence proteins of H. pylori among quantitatively meaningful ten spots and confirmed the 2-DE results by Western blot analysis. These results suggest that cyanidin 3-O-glucoside can modulate a variety of H. pylori pathogenic determinants.

Effect of High-Temperature Post-Oxidation Annealing in Diluted Nitric Oxide Gas on the SiO2/4H-SiC Interface (4H-SiC와 산화막 계면에 대한 혼합된 일산화질소 가스를 이용한 산화 후속 열처리 효과)

  • In kyu Kim;Jeong Hyun Moon
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.37 no.1
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    • pp.101-105
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    • 2024
  • 4H-SiC power metal-oxide-semiconductor field effect transistors (MOSFETs) have been developed to achieve lower specific-on-resistance (Ron,sp), and the gate oxides have been thermally grown. The poor channel mobility resulting from the high interface trap density (Dit) at the SiO2/4H-SiC interface significantly affects the higher switching loss of the power device. Therefore, the development of novel fabrication processes to enhance the quality of the SiO2/4H-SiC interface is required. In this paper, NO post-oxidation annealing (POA) by using the conditions of N2 diluted NO at a high temperature (1,300℃) is proposed to reduce the high interface trap density resulting from thermal oxidation. The NO POA is carried out in various NO ambient (0, 10, 50, and 100% NO mixed with 100, 90, 50, and 0% of high purity N2 gas to achieve the optimized condition while maintaining a high temperature (1,300℃). To confirm the optimized condition of the NO POA, measuring capacitance-voltage (C-V) and current-voltage (I-V), and time-of-flight secondary-ion mass spectrometry (ToF-SIMS) are employed. It is confirmed that the POA condition of 50% NO at 1,300℃ facilitates the equilibrium state of both the oxidation and nitridation at the SiO2/4H-SiC interface, thereby reducing the Dit.

Multicomponent assessment and ginsenoside conversions of Panax quinquefolium L. roots before and after steaming by HPLC-MSn

  • Huang, Xin;Liu, Yan;Zhang, Yong;Li, Shuai-Ping;Yue, Hao;Chen, Chang-Bao;Liu, Shu-Ying
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.27-37
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    • 2019
  • Background: The structural conversions in ginsenosides induced by steaming or heating or acidic condition could improve red ginseng bioactivities significantly. In this paper, the chemical transformations of red American ginseng from fresh Panax quinquefolium L. under steaming were investigated, and the possible mechanisms were discussed. Methods: A method with reversed-phase high-performance liquid chromatography coupled with linear ion trap mass spectrometry ($HPLC-MS^n$)-equipped electrospray ionization ion source was developed for structural analysis and quantitation of ginsenosides in dried and red American ginseng. Results: In total, 59 ginsenosides of protopanaxadiol, protopanaxatriol, oleanane, and ocotillol types were identified in American ginseng before and after steaming process by matching the molecular weight and/or comparing $MS^n$ fragmentation with that of standards and/or known published compounds, and some of them were determined to be disappeared or newly generated under different steaming time and temperature. The specific fragments of each aglycone-type ginsenosides were determined as well as aglycone hydrated and dehydrated ones. The mechanisms were deduced as hydrolysis, hydration, dehydration, and isomerization of neutral and acidic ginsenosides. Furthermore, the relative peak areas of detected compounds were calculated based on peak areas ratio. Conclusion: The multicomponent assessment of American ginseng was conducted by $HPLC-MS^n$. The result is expected to provide possibility for holistic evaluation of the processing procedures of red American ginseng and a scientific basis for the usage of American ginseng in prescription.

Differential Expression of O-glycoprotein Glycans in Cholangiocarcinoma Cell Lines

  • Talabnin, Krajang;Talabnin, Chutima;Ishihara, Mayumi;Azadi, Parastoo;Wongkham, Sopit;Sripa, Banchob
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.691-695
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    • 2016
  • Protein glycosylation is the most common posttranslational modification in mammalian cells. Aberrant protein glycosylation has been reported in various diseases, including cancer. We identified and quantified the glycan structures of O-linked glycoprotein from cholangiocarcinoma (CCA) cell lines from different histological types and compared their profiles by nanospray ionization-linear ion trap mass spectrometry (NSI-$MS^n$). Five human CCA cell lines, K100, M055, M139, M213 and M214 were characterized. The results showed that the O-linked glycans of the CCA cell lines comprised tri- to hexa-saccharides with terminal galactose and sialic acids: NeuAc1Gal1GalNAc1, Gal2GlcNAc1GalNAc1, NeuAc2Gal1GalNAc1 NeuAc1Gal2GlcNAc1GalNAc1 and NeuAc2Gal2GlcNAc1GalNAc1 All five CCA cell lines showed a similar glycan pattern, but with differences in their quantities. NeuAc1Gal1GalNAc1 proved to be the most abundant structure in poorly differentiated adenocarcinoma (K100; 57.1%), moderately differentiated adenocarcinoma (M055; 42.6%) and squamous cell carcinoma (M139; 43.0%), while moderately to poorly differentiated adenocarcinoma (M214; 40.1%) and adenosquamous cell carcinoma (M213; 34.7%) appeared dominated by $NeuA_{c2}Gal_1GalNA_{c1}$. These results demonstrate differential expression of the O-linked glycans in the different histological types of CCA. All five CCA cell lines have abundant terminal sialic acid (NeuAc) O-linked glycans, suggesting an important role for sialic acid in cancer cells. Our structural analyses of glycans may provide important information regarding physiology of disease-related glycoproteins in CCA.

Quantitative Determination of PFOA and PFOS in the Effluent of Sewage Treatment Plants and in Han River (서울시 하수처리장 방류수 및 한강 내 PFOA와 PFOS의 과불화화합물 모니터링 연구)

  • Shin, Mi-Yeon;Im, Jong-Kwon;Kho, Young-Lim;Choi, Kyoung-Sik;Zoh, Kyung-Duk
    • Journal of Environmental Health Sciences
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    • v.35 no.4
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    • pp.334-342
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    • 2009
  • Perfluorinated compounds (PFCs) have a wide range of domestic and industrial applications, but they are persistent in the environment. Perfluorooctanoic acid (PFOA) and perfluorooctanesulfonate (PFOS) are among the metabolites of PFCs and occur at high concentration in the environment. Korea is the largest importer of PFC compounds in the world, therefore, the accumulation of these compounds is possible. In this study, the concentrations of PFOS and PFOA were determined in water samples taken from sewage treatment plants (STPs) and the Han River in Seoul, Korea. After extraction with a HLB cartridge, PFCs in the samples were analyzed by HPLC with an ion trap mass spectrometry in electrospray negative mode. Limits of detection was between 1 and 1.6 ng/l. The result showed that the concentrations of PFOS and PFOA in effluent and influent of the four STPs in Seoul were 60~570 ng/l, and not detected (nd)~254 ng/l, respectively. The levels of PFOS and PFOA were higher in the effluents which passed through the treatment process than in influent water samples which was against expectation. The concentration of PFOA and PFOS in the Han River was 60~570 ng/l and nd~254 ng/l, respectively. PFOA was detected in every sample, but PFOS was only detected in the downstreams of the Han River. This result indicates that there is comprehensive contamination of PFCs in the aquatic environment in Korea.

Quercetin Glucoside Profiling of Fresh Onion (Allium cepa) and Aged Black Onion Using HPLC-ESI/MS/MS (HPLC-ESI/MS/MS를 이용한 생양파와 흑양파의 퀘세틴 배당체 분석)

  • Chung, Dong-Min;Kwon, Sun-Hwa;Chung, Young-Chul;Chun, Hyo-Kon
    • Journal of Life Science
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    • v.21 no.3
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    • pp.464-467
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    • 2011
  • Quercetin is a major flavonoid present in onions, which acts as an antioxidant. Quercetin exists both as a free compound and conjugated with carbohydrates, primarily as glucosides in onion. Aged black onion was made through a 30 day aging process in which the onions were kept in an environment of $60^{\circ}C$ and high humidity (90% RH). Quercetin and quercetin glucosides were assayed in onion bulbs before and after the aging process, using high performance liquid chromatography-electrospray ion trap mass spectrometry (HPLC-ESI/MS/MS). Quercetin mono- and diglucosides were identified in fresh onion bulbs, whereas quercetin aglycone was the only form present in aged black onion bulbs. These findings indicate that the quercetin mono- and di-glucosides present in fresh onions undergo complete deglycosylation during the aging process. Such profiling will provide a rapid method that can be used to assess changes in the two major quercetin glycosides during the aging process of onion bulbs.

Transcriptome Analysis Reveals the Putative Polyketide Synthase Gene Involved in Hispidin Biosynthesis in Sanghuangporus sanghuang

  • Jiansheng Wei;Liangyan Liu;Xiaolong Yuan;Dong Wang;Xinyue Wang;Wei Bi;Yan Yang;Yi Wang
    • Mycobiology
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    • v.51 no.5
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    • pp.360-371
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    • 2023
  • Hispidin is an important styrylpyrone produced by Sanghuangporus sanghuang. To analyze hispidin biosynthesis in S. sanghuang, the transcriptomes of hispidin-producing and non-producing S. sanghuang were determined by Illumina sequencing. Five PKSs were identified using genome annotation. Comparative analysis with the reference transcriptome showed that two PKSs (ShPKS3 and ShPKS4) had low expression levels in four types of media. The gene expression pattern of only ShPKS1 was consistent with the yield variation of hispidin. The combined analyses of gene expression with qPCR and hispidin detection by liquid chromatography-mass spectrometry coupled with ion-trap and time-of-flight technologies (LCMS-IT-TOF) showed that ShPKS1 was involved in hispidin biosynthesis in S. sanghuang. ShPKS1 is a partially reducing PKS gene with extra AMP and ACP domains before the KS domain. The domain architecture of ShPKS1 was AMP-ACP-KS-AT-DH-KR-ACP-ACP. Phylogenetic analysis shows that ShPKS1 and other PKS genes from Hymenochaetaceae form a unique monophyletic clade closely related to the clade containing Agaricales hispidin synthase. Taken together, our data indicate that ShPKS1 is a novel PKS of S. sanghuang involved in hispidin biosynthesis.