• Title/Summary/Keyword: Laser mass spectrometry

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Proteome Analysis of Vernalization-Treated Arabidopsis thaliana by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Cho, Mi-Ran;Lee, Kyung-Hyeon;Hyun, You-Bong;Lee, Il-Ha;Kim, Hie-Joon
    • Bulletin of the Korean Chemical Society
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    • v.28 no.3
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    • pp.427-431
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    • 2007
  • In order to gain insight into the molecular changes at the protein level in plants exposed to low temperature for a long period of time (vernalization), proteome analyses of vernalization-treated Arabidopsis thaliana have been carried out by two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. Fourteen proteins including ATP binding/GTP binding/translation elongation factor and glycine-rich RNA-binding protein 7 (GRP7) showed differential expression in vernalization-treated Arabidopsis thaliana. GRP7 showed the most dramatic increase in expression suggesting its involvement in response to vernalization treatment.

Fast visible dye staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gels compatible with matrix-assisted laser desorption/ionization-mass spectrometry

  • Jin, Li-Tai;Hwang, Sun-Young;Yoo, Gyurng-Soo;Choi, Jung-Kap
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.147.2-147.2
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    • 2003
  • A fast and matrix-assisted laser desorption/ionization-mass spectrometry compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, Zincon and Ethyl Violet to form an ion-pair complex. It is safe to use since the methanol used previously in staining solution was replaced with ethanol, which is not toxic. The protocol including fixing, staining and quick washing steps can be completed in 1 to 1.5 h depending upon gel thickness. (omitted)

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Microwave-assisted Weak Acid Hydrolysis of Proteins

  • Seo, Mi-Yeong;Kim, Jin-Hee;Park, Se-Hwan;Lee, Ji-Hye;Kim, Tae-Hee;Lee, Ji-Hyeon;Kim, Jeong-Kwon
    • Mass Spectrometry Letters
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    • v.3 no.2
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    • pp.47-49
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    • 2012
  • Myoglobin was hydrolyzed by microwave-assisted weak acid hydrolysis with 2% formic acid at $37^{\circ}C$, $50^{\circ}C$, and $100^{\circ}C$ for 1 h. The most effective hydrolysis was observed at $100^{\circ}C$. Hydrolysis products were investigated using matrixassisted laser desorption/ionization time-of-flight mass spectrometry. Most cleavages predominantly occurred at the C-termini of aspartyl residues. For comparison, weak acid hydrolysis was also performed in boiling water for 20, 40, 60, and 120 min. A 60- min weak acid hydrolysis in boiling water yielded similar results as a 60-min microwave-assisted weak acid hydrolysis at $100^{\circ}C$. These results strongly suggest that microwave irradiation has no notable enhancement effect on acid hydrolysis of proteins and that temperature is the major factor that determines the effectiveness of weak acid hydrolysis.

Identification of antigenic proteins of lymphocystis disease virus (LCDV) by MALDI-TOF mass spectrometry

  • Chung, Chang-Kyun;Kim, Byung-Gwan;Jung, Myung-Hwa;Jung, Sung-Ju
    • Journal of fish pathology
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    • v.28 no.3
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    • pp.133-143
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    • 2015
  • The antigenic proteins of Lymphocystis disease virus (LCDV) from tumors of olive flounder, Paralichthys olivaceus, are described following characterization by mass spectrometry. In SDS-PAGE, predominant protein bands were observed at 114, 88, 70, 54, 52, 47, 42 and 24 kDa. Western blot analysis showed that antisera reacted strongly at molecular weights of 114, 67 and 54 kDa, and reacted weakly at molecular weights of 74, 70, 36, 24 and 22 kDa. In the identification of LCDV antigenic proteins by matrix-assisted laser desorption ionization (MALDI) TOF mass spectrometry, 10 of 14 excised bands consisted mostly of proteins with amino acid sequences that matched LCDV-C (lymphocystis disease virus isolate China) ORFs. Strong antigens with molecular weights of 114, 67 and 54 kDa were identified as LDVICp236 (chromosome segregation ATPase), LDVICp033 (membrane bound metallopeptidase) and LDVICp157 (hypothetical protein), respectively. Minor antigens with molecular weights of 70, 36, 24 and 22 kDa proteins were identified as LDVICp160 (acetyl-coA hydrolase), LDVICp213 (hypothetical protein), LDVICp039 (hypothetical protein) and LDVICp213 (hypothetical protein). However, the major capsid protein (LDVICp043) did not react with the polyclonal antibody.

Comparison of Matrices for Optimal Analysis of Synthetic Polymers Using MALDI-TOF Mass Spectrometry

  • Yoo, Hee-Jin;Kim, Duck-Hyun;Choi, Yoon-Ji;Choi, Jung Hoon;Park, Moonhee;Shin, DongJin;Oh, YoonSeok;Kim, YangSun;Cho, Kun
    • Mass Spectrometry Letters
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    • v.11 no.4
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    • pp.77-81
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    • 2020
  • Characterization of the various chemical aspects of composite polymers is important for quality control of manufactured polymers. In this study, we compared three suitable matrices (α cyano-4-hydroxycinnamic acid [CHCA], 2,5 dihydroxy benzoic acid [2,5-DHB], and dithranol), to characterize various synthetic polymers by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Although the spectra obtained with the CHCA and 2,5-DHB matrices were generally good, in certain samples ghost peaks disappeared only when dithranol was used as the matrix. Furthermore, we examined the use of sodium trifluoroacetate (NaTFA) as an additive to reduce interference by metals and copolymers in the spectra. In conclusion, appropriate selection of a matrix, according to the characteristics of the polymer, and the use of additives to improve sensitivity are important considerations for polymer analysis and development.

Discrimination of Bacillus anthracis Spores by Direct in-situ Analysis of Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry

  • Jeong, Young-Su;Lee, Jonghee;Kim, Seong-Joo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2635-2639
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    • 2013
  • The rapid and accurate identification of biological agents is a critical step in the case of bio-terror and biological warfare attacks. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been widely used for the identification of microorganisms. In this study, we describe a method for the rapid and accurate discrimination of Bacillus anthracis spores using MALDI-TOF MS. Our direct in-situ analysis of MALDI-TOF MS does not involve subsequent high-resolution mass analyses and sample preparation steps. This method allowed the detection of species-specific biomarkers from each Bacillus spores. Especially, B. anthracis spores had specific biomarker peaks at 2503, 3089, 3376, 6684, 6698, 6753, and 6840 m/z. Cluster and PCA analyses of the mass spectra of Bacillus spores revealed distinctively separated clusters and within-groups similarity. Therefore, we believe that this method is effective in the real-time identification of biological warfare agents such as B. anthracis as well as other microorganisms in the field.

Mass Spectrometric Study of Carbon Cluster Formation in Laser Ablation of Graphite at 355 nm

  • Koo, Young-Mi;Choi, Young-Ku;Lee, Kee-Hag;Jung, Kwang-Woo
    • Bulletin of the Korean Chemical Society
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    • v.23 no.2
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    • pp.309-314
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    • 2002
  • The ablation dynamics and cluster formation of $C_n^+$ ions ejected from 355 nm laser ablation of a graphite target in vacuum are investigated using a reflectron time-of-flight (RTOF) mass spectrometer. At low laser fluence, odd-numbered cluster ions with $3{\leq}n{\leq}15$ are predominantly produced. Increasing the laser fluence shifts the maximum size distribution towards small cluster ions, implying the fragmentation of larger clusters within the hot plume. The temporal evolution of $C_n^+$ ions was measured by varying the delay time of the ion extraction pulse with respect to the laser irradiation, providing significant information on the characteristics of the ablated plume. Above a laser fluence of $0.2J/cm^2$ , large cluster ions ($n{\geq}30$) are produced at relatively long delay times, indicating that atoms or small carbon clusters aggregate during plume propagation. The dependence of the intensity of ablated $C_n^+$ ions on delay time after laser irradiation shows that the most probable velocity of each cluster ion decreases with cluster size.

Selective or Class-wide Mass Fingerprinting of Phosphatidylcholines and Cerebrosides from Lipid Mixtures by MALDI Mass Spectrometry

  • Lee, Gwangbin;Son, Jeongjin;Cha, Sangwon
    • Bulletin of the Korean Chemical Society
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    • v.34 no.7
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    • pp.2143-2147
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    • 2013
  • Matrix assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is a very effective method for lipid mass fingerprinting. However, MALDI MS suffered from spectral complexities, differential ionization efficiencies, and poor reproducibility when analyzing complex lipid mixtures without prior separation steps. Here, we aimed to find optimal MALDI sample preparation methods which enable selective or class-wide mass fingerprinting of two totally different lipid classes. In order to achieve this, various matrices with additives were tested against the mixture of phosphatidylcholine (PC) and cerebrosides (Cers) which are abundant in animal brain tissues and also of great interests in disease biology. Our results showed that, from complex lipid mixtures, 2,4,6-trihydroxyacetophenone (THAP) with $NaNO_3$ was a useful MALDI matrix for the class-wide fingerprinting of PC and Cers. In contrast, THAP efficiently generated PC-focused profiles and graphene oxide (GO) with $NaNO_3$ provided Cer-only profiles with reduced spectral complexity.