• Title/Summary/Keyword: matrix assisted laser desorption ionization mass spectrometry

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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.

Comparative Analysis of Serum Proteomes of Moyamoya Disease and Normal Controls

  • Koh, Eun-Jeong;Kim, Han-Na;Ma, Tian-Ze;Choi, Ha-Young;Kwak, Yong-Geun
    • Journal of Korean Neurosurgical Society
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    • v.48 no.1
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    • pp.8-13
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    • 2010
  • Objective : The etiology and pathogenesis of moyamoya disease remain unclear. Furthermore, the definitive diagnostic protein-biomarkers for moyamoya disease are still unknown. The present study analyzed serum proteomes from normal controls and moyamoya patients to identify novel serological biomarkers for diagnosing moyamoya disease. Methods : We compared the two-dimensional electrophoresis patterns of sera from moyamoya disease patients and normal controls and identified the differentially-expressed spots by matrix-assisted laser desorption/ionization-time-of flight mass spectrometry and electrospray ionization quadruple time-of-flight mass spectrometry. Results : We found and analyzed 22 differently-expressed proteomes. Two proteins were up-regulated. Twenty proteins were down-regulated. Complement C1 inhibitor protein and apolipoprotein C-III showed predominantly changed expressions (complement C1 inhibitor protein averaged a 7.23-fold expression in moyamoya patients as compared to controls, while apolipoprotein C-III averaged a 0.066-fold expression). Conclusion : Although our study had a small sample size, our proteomic data provide serologic clue proteins for understanding moyamoya disease.

Characterization of Yeast and Bacterial Type Strains with Food and Agricultural Applications by MALDI-TOF Mass Spectrometry Biotyping

  • Harnpicharnchai, Piyanun;Jaresitthikunchai, Janthima;Seesang, Mintra;Jindamorakot, Sasitorn;Tanapongpipat, Sutipa;Ingsriswang, Supawadee
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.138-147
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    • 2020
  • Various microorganisms play important roles in food fermentation, food spoilage, and agriculture. In this study, the biotype of 54 yeast and bacterial strains having high potential for utilization in food and agriculture, including Candida spp., Lactobacillus spp., and Acetobacter spp., were characterized by matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS). This characterization using a fast and robust method provides much-needed information on the selected microorganisms and will facilitate effective usage of these strains in various applications. Importantly, the unique protein profile of each microbial species obtained from this study was used to create a database of fingerprints from these species. The database was validated using microbial strains of the same species by comparing the mass spectra with the created database through pattern matching. The created reference database provides crucial information and is useful for further utilization of a large number of valuable microorganisms relevant to food and agriculture.

Effect of Dihydroxybenzoic Acid Isomers on the Analysis of Polyethylene Glycols in MALDI-MS

  • Lee, Ae-Ra;Yang, Hyo-Jik;Kim, Yang-Sun;Kim, Jeong-Kwon
    • Bulletin of the Korean Chemical Society
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    • v.30 no.5
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    • pp.1127-1130
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    • 2009
  • The effects of different dihydroxybenzoic acid (DHB) isomers, when used as matrix materials in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), were investigated in analyses of polyethylene glycol (PEG) polymers. PEG polymers ranging from 400 to 8,000 Da were prepared in different DHB isomer matrices using solvent-based and solvent-free methods. PEG samples were detected only in matrices of 2,3-DHB, 2,5-DHB, and 2,6-DHB while the most intense peaks were observed using 2,6-DHB in both solvent-free and solvent-based preparations.

Application of MALDI Tissue Imaging of Drugs and Metabolites: A New Frontier for Molecular Histology

  • Shanta, Selina Rahman;Kim, Young-Jun;Kim, Young-Hwan;Kim, Kwang-Pyo
    • Biomolecules & Therapeutics
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    • v.19 no.2
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    • pp.149-154
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    • 2011
  • Matrix assisted laser desorption ionization (MALDI) mass spectrometry is commonly used to analyze biological molecules such as proteins, peptides and lipids from cells or tissue. Recently MALDI Imaging mass spectrometry (IMS) has been widely applied for the identification of different drugs and their metabolites in tissue. This special feature has made MALDI-MS a common choice for investigation of the molecular histology of pathological samples as well as an important alternative to other conventional imaging methods. The basic advantages of MALDI-IMS are its simple technique, rapid acquisition, increased sensitivity and most prominently, its capacity for direct tissue analysis without prior sample preparation. Moreover, with ms/ms analysis, it is possible to acquire structural information of known or unknown analytes directly from tissue sections. In recent years, MALDI-IMS has made enormous advances in the pathological field. Indeed, it is now possible to identify various changes in biological components due to disease states directly on tissue as well as to analyze the effect of treated drugs. In this review, we focus on the advantages of MALDI tissue imaging over traditional methods and highlight some motivating findings that are significant in pathological studies.

Comparative Proteome Analysis of Celastrol-Treated Helicobacter pylori

  • Kim, Sa-Hyun
    • Biomedical Science Letters
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    • v.23 no.4
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    • pp.395-401
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    • 2017
  • Various preclinical and clinical trials have been conducted the efficacy of celastrol. In data presented in the current manuscript is the first trial to inhibit Helicobacter pylori with celastrol. In this study, the quantitative change of various H. pylori proteins including CagA and VacA by the anti-bacterial effect of celastrol was determined. The anti-H. pylori effects of celastrol was investigated by performing 2-dimensional electrophoresis and additional supporting experiments. After 2-dimensional electrophoresis analysis, spot intensities were analyzed and 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). The results show that celastrol has multiple effects on protein expression in H. pylori.

Postharvest Biological Control of Colletotrichum acutatum on Apple by Bacillus subtilis HM1 and the Structural Identification of Antagonists

  • Kim, Hae-Min;Lee, Kui-Jae;Chae, Jong-Chan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1954-1959
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    • 2015
  • Bacillus subtilis HM1 was isolated from the rhizosphere region of halophytes for its antifungal activity against Colletotrichum acutatum, the causative agent of anthracnose. Treatment of postharvest apples with the cell culture or with a cell-free culture supernatant reduced disease severity 80.7% and 69.4%, respectively. Both treatments also exhibited antifungal activity against various phytopathogenic fungi in vitro. The antifungal substances were purified and analyzed by acid precipitation, gel filtration, high-performance liquid chromatography, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Three compounds were identified as fengycin, iturin, and surfactin. The MALDI-TOF/TOF mass spectrum revealed the presence of cyclized fengycin homologs A and B, which were distinguishable on the basis of the presence of either alanine or valine, respectively, at position 6 of the peptide sequence. In addition, the cyclized structure of fengycin was shown to play a critical role in antifungal activity.

Selective Extraction and Quantification of Glutathione using Maleimide-Presenting Gold Nanoparticles

  • Oh, Hongseok;Lee, Jeongwook;Yeo, Woon-Seok
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.3047-3051
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    • 2014
  • In this paper, we describe a new method for the selective extraction and quantification of glutathione (GSH) using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and maleimide-presenting gold nanoparticles (Mal-AuNPs). Our strategy utilizes the Michael addition to selectively extract GSH, from chosen samples, onto the maleimide of Mal-AuNPs. After the extraction step, the GSH bound to the AuNPs was analyzed by MALDI-TOF MS in the presence of an internal standard which was prepared by reacting Mal-AuNPs with isotope-labeled GSH ($GSH^*$). The $GSH^*$ has the same structure as GSH but a higher molecular weight, and therefore, enables absolute quantification of GSH by comparing the mass signal intensities of the GSH- and $GSH^*$-conjugated alkanethiols. Our strategy was verified by analyzing GSH-spiked fetal bovine serum and NIH 3T3 cells.

Characterization of Thermal Degradation of Polytrimethylene Terephthalate by MALDI-TOF Mass Spectrometry

  • Jang, Sung-Woo;Yang, Eun-Kyung;Jin, Sung-Il;Cho, Young-Dal;Choe, Eun-Kyung;Park, Chan-Ryang
    • Bulletin of the Korean Chemical Society
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    • v.33 no.3
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    • pp.833-838
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    • 2012
  • The thermal degradation products of polytrimethylene terephthalate (PTT) obtained by heating the sample in the temperature range of $250-360^{\circ}C$ under non-oxidative conditions was characterized using MALDI-TOF (matrix assisted laser desorption/ionization) mass spectrometry. The structures of the degradation products were determined and the relative compositions were estimated. The MALDI-TOF mass spectra of the thermally degraded PTT sample showed three main series of oligomer products with different end groups, which were carboxyl/carboxyl, carboxyl/allyl, and allyl/allyl. In contrast to the thermal degradation of polyethylene terephthalate (PET), the oligomers containing terephthalic anhydrides were not detected, whereas the formation of oligomers containing the unsaturated allyl ester group was confirmed by mass assignment. From these results, it was concluded that the thermal degradation of PTT proceeds exclusively through the ${\beta}$-CH hydrogen transfer mechanism, which is in accordance with the proposed reaction mechanism for the thermal degradation of polybutylene terephthalate (PBT).

Study on the variation of cellular physiology of Escherichia coli during high cell density cultivation using 2-dimensional gel electrophoresis

  • Yun, Sang-Seon;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.219-222
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    • 2000
  • Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

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