• Mass Spectrometry Letters
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• v.12 no.3
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• pp.81-84
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• 2021

Collagen, which accounts for one-third of human protein, is reduced due to human aging, and much attention is focused on making collagen into food to prevent such aging. Gel permeation chromatography with Reflective Index (RI) detection (GPC/RI) was chosen as the most suitable instrument to confirm molecular weight distribution, and we explored the use of this technique for analysis of collagen peptide molecular sizes and distributions. Data reliability was verified by matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometric analysis. The data were considered meaningful for comparative analysis of molecular weight distribution patterns.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.3
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• pp.275-283
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• 2018

Helicobacter pylori is a causative organism of various gastrointestinal diseases, including chronic gastritis, gastric ulcer, or gastric adenocarcinoma. Pathogenic factors, such as cytotoxin-associated protein A (CagA) and vacuolating cytotoxic protein A (VacA), play a role. This study analyzed qualitatively and quantitatively the effects of zerumbone on the changes in the protein expression levels of various H. pylori proteins, including CagA and VacA. Approximately 200 significant proteins were screened for the H. pylori 60190 (VacA positive / CagA positive; Eastern type) strain, and proteomic analysis was performed on 13 protein molecules that were clinically significant. After two-dimensional electrophoresis (2-DE), $ImageMaster^{TM}$ 2-DE Platinum software was used for quantitative measurements of protein spots. Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-TOF-MS) and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) were used for protein identification. After intensive analysis of the proteins that showed significant changes, a reverse transcription-polymerase chain reaction was performed as required to verify the results. In this study, the significance of zerumbone as a therapeutic agent for H. pylori infection was examined by screening a new pharmacological activity mechanism of zerumbone.

• Mass Spectrometry Letters
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• v.2 no.3
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• pp.61-64
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• 2011

Matrix Assisted Laser Desorption/Ionization-Time-of-Flight mass spectrometry (MALDI-TOF-MS) is the most widely used MS technique for glycan analysis. However, the poor point-to-point and sample-to-sample reproducibility becomes a limit in glycan biomarker research. A prespotted MALDI plate which overcomes the large crystal formation of 2,5-dihydroxybenzoic acid (DHB) has been developed and applied for glycan analysis. A homogeneous matrix coated surface without a crystal structure was formed on a hydrophilic/ hydrophobic patterned surface using a piezoelectric device. The reproducible MALDI-TOF-MS data have been presented using MALDI imaging of beer glycan as well as serum glycan eluted from 10% and 20% ACN elution fractions. The glycan profile from the serum glycan by MALDI-TOF-MS with a DHB prespotted plate was highly conserved for 10 different spectra and the coefficient of variations of significant ion peaks of MALDI data varies from 3.59 to 19.95.

• Molecules and Cells
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• v.24 no.2
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• pp.268-275
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• 2007

Protein phosphorylation plays a key role in signal transduction in cells. Since phosphoproteins are present in low abundance, enrichment methods are required for their purification and analysis. Chemical derivatization strategies have been devised for enriching phosphoproteins and phosphopeptides. In this report, we employed a strategy that replaces the phosphate moieties on serine and threonine residues with a biotin-containing tag via a series of chemical reactions. Ribulose 1,5-bisphosphate carboxylase/oxygenase (RUBISCO)-depleted protein extracts prepared from Arabidopsis seedlings were chemically modified for 'biotin-tagging'. The biotinylated (previously phosphorylated) proteins were then selectively isolated by avidin-biotin affinity chromatography, followed by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). This led to the identification of 31 protein spots, representing 18 different proteins, which are implicated in a variety of cellular processes. Despite its current technical limitations, with further improvements in tools and techniques this strategy may be developed into a useful approach.

• Journal of Microbiology and Biotechnology
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• v.17 no.2
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• pp.207-217
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• 2007

The Saccharomyces0 cerevisiae KNU5377 strain, which was isolated from spoilage in nature, has the ability to convert biomass to alcohol at high temperatures and it can resist against various stresses [18, 19]. In order to understand the defense mechanisms of the KNU5377 strain under menadione (MD) as oxidative stress, we used several techniques for study: peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) followed by two-dimensional (2D) gel electrophoresis, liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), and surface-enhanced laser desorption ionization-time of flight (SELDI-TOF) technology. Among the 35 proteins identified by MALDI-TOF MS, 19 proteins including Sod1p, Sod2p, Tsa1p, and Ahp1p were induced under stress condition, while 16 proteins were augmented under normal condition. In particular, five proteins, Sod1p, Sod2p, Ahp1p, Rib3p, Yaf9p, and Mnt1p, were induced in only stressed cells. By LC-ESI-MS/MS analysis, 37 proteins were identified in normal cells and 49 proteins were confirmed in the stressed cells. Among the identified proteins, 32 proteins were found in both cells. Five proteins including Yel047cp and Met6p were only upregulated in the normal cells, whereas 17 proteins including Abp1P and Sam1p were elevated in the stressed cells. It was interesting that highly hypothetical proteins such as Ynl281wp, Ygr279cp, Ypl273wp, Ykl133cp, and Ykr074wp were only expressed in the stressed cells. SELDI-TOF analysis using the SAX2 and WCX2 chips showed that highly multiple-specific protein patterns were reproducibly detected in ranges from 2.9 to 27.0 kDa both under normal and stress conditions. Therefore, induction of antioxidant proteins, hypothetical proteins, and low molecular weight proteins were revealed by different proteomic techniques. These results suggest that comparative analyses using proteomics might contribute to elucidate the defense mechanisms of KNU5377 under MD stress.

• Bulletin of the Korean Chemical Society
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• v.25 no.12
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• pp.1791-1800
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• 2004

Angiotensin I, a model decapeptide, was glycosylated and partially hydrolyzed with HCl (6 N, 80 $^{\circ}C$, 4 h), aminopeptidase, and carboxypeptidase Y. A single peptide mass map obtained from truncated peptides in the partial acid hydrolysate of angiotensin and its glycosylation product mixture by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry enabled sequencing of angiotensin by a combinatorial procedure. MALDI-TOF and electrospray ionization (ESI) tandem mass spectrometric results indicate that both the N-terminal amino group of aspartic acid and the guanidinium group of the second residue arginine are glycosylated.

• Korean Journal of Veterinary Research
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• v.44 no.1
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• pp.57-64
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• 2004

The common pathogen Salmonella enteirtidis (S. enteritidis) is the major cause of foodborne disease. Protein identification by peptide mass fingerprinting using the matrix-assisted laser desorption ionization time of fight (MALDI-TOF) mass spectrometry (MS) can analysis unambiguously identity the spots from 2-dimensional electrophoresis (2-DE) gel. In this report, we examined protein components from patterns of S. enteritidis proteins. In addition, antigens that are recognized by sera can be identified by immunoblotting. This study that 2-DE analysis of S. enteritidis yields useful information concerning S. enteritidis proteome, the results that have been obtained led to a more detailed understanding of Salmonella pathology and open further interesting fields for future work.

• Journal of Microbiology and Biotechnology
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• v.16 no.1
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• pp.25-31
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• 2006

Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

• 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.

• Macromolecular Research
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• v.16 no.2
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• pp.108-112
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• 2008

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) analysis of nonpolar polymeric materials is affected by the sample preparation as well as the matrix and cationizing agent. This study examined the influence of silver salt types on the MALDI analysis of polybutadiene (PB). Silver trifluoroacetate (AgTFA), silver benzoate (AgBz), silver nitrate ($AgNO_3$), and silver p-toluenesulfonate (AgTS) were used as the silver salts to compare the MALDI mass spectra of PB. The mixture solution of PB and 2,5-dihydroxybenzoic acid (DHB), as a matrix dissolved in THF, was spotted on the sample plate and dried. A droplet of the aqueous silver salt solution was placed onto the mixture. The mass spectrum with AgBz showed the clear $[M+Ag]^+$ ion distribution of PB while the mass spectrum with AgTFA did not show $[M+Ag]^+$ ions but only silver cluster ions. The mass spectra with $AgNO_3$ and AgTS did not show a clear $[M+Ag]^+$ ion distribution. The difference in the formation of $[M+Ag]^+$ ions of PB depending on the silver salts was attributed to the silver cation transfer reaction between the silver salt and the matrix (DHB). The mass spectrum showed a clear $[M+Ag]^+$ ion distribution of PB when the conjugate acid of the silver salt was less acidic than the matrix.