• Mass Spectrometry Letters
• /
• 제10권3호
• /
• pp.84-87
• /
• 2019

Liquid chromatography mass spectrometry is widely employed in proteomics studies. One of such instruments is the Liquid Chromatography (LC)-Matrix-assisted laser desorption ionisation (MALDI)-Time of flight (TOF) or LC-MALDI-TOF/TOF. In this study, this instrument was used to identify the membrane proteins of a protozoan parasite namely Entamoeba histolytica. It causes amoebiasis in human. The E. histolytica trophozoites were cultured prior to the membrane protein extraction using the conventional method, $ProteoPrep^{(R)}$ and $ProteoExtract^{(R)}$ kits. Then, the membrane protein extracts were trypticdigested and analysed by LC-MALDI-TOF/TOF. Approximately, 194 proteins were identified and 27.8% (54) were predicted as membrane proteins having 1 to 15 transmembrane regions and signal peptides by combining all three extraction methods. Also, this study has discovered 3 unique proteins as compared to our previous study which merit further investigation.

• Bulletin of the Korean Chemical Society
• /
• 제31권10호
• /
• pp.2791-2796
• /
• 2010

In the present study, we constructed a membrane inlet assembly for selective permeation of volatile airborne organic compounds for subsequent analysis by time-of-flight mass spectrometry. The time-dependent diffusion of analytes through a $75\;{\mu}m$ thick polydimethylsiloxane membrane was measured by monitoring the ion signal after a step change in the sample concentration. The results fit well to a non-steady-state permeation equation. The diffusion coefficient, response time, and sensitivity were determined experimentally for a range of polar (halogenated) and nonpolar (aromatic) compounds. We found that the response times for several volatile organic compounds were greatly influenced by the alkyl chain length as well as the size of the substituted halogen atoms. The detection limits for benzene, ethylbenzene, and 2-propanol were 0.2 ppm, 0.1 ppm, and 3.0 ppm by volume, respectively, with a linear dynamic range greater than three orders of magnitude. These results indicate that the membrane inlet/time-of-flight mass spectrometry technique will be useful for a wide range of applications, particularly for in situ environmental monitoring.

• Mass Spectrometry Letters
• /
• 제11권2호
• /
• pp.25-29
• /
• 2020

Pseudopodia are dynamic actin cytoskeleton-based membrane protrusions of cells that enable directional cell migration. Pseudopodia of cancer cells play key roles in cancer metastasis. Recent studies using pseudopodial subcellular fractionation methodologies combined with mass spectrometry-based proteomic profiling have provided insight into the pseudopodiome that control the protrusions of invasive metastatic cancer cells. This review highlights how to characterize the protein composition of pseudopodia and develop strategies to identify biomarkers or drug candidates that target reduction or prevention of metastatic cancer.

• Mass Spectrometry Letters
• /
• 제9권3호
• /
• pp.67-72
• /
• 2018

Alkaline phosphatase (AP) is a membrane-bound glycoprotein that is widely distributed in the plasma membrane of cells of various organs and also found in many organisms from bacteria to humans. The complete amino acid sequence and three-dimensional structure of human placental alkaline phosphatase have been reported. Based on the literature data, AP consists of two presumptive glycosylation sites, at Asn-144 and Asn-271. However, it only contains a single occupied N-linked glycosylation site and no occupied O-linked glycosylation sites. Hydrophilic interaction chromatography (HILIC) has been primarily employed for the characterization of the glycan structures derived from glycoproteins. N-glycan structures from human placental alkaline phosphatase (PLAP) were investigated using HILIC-Orbitrap MS, and subsequent data processing and glycan assignment software. 16 structures including 10 sialylated N-glycans were identified from PLAP.

• Mass Spectrometry Letters
• /
• 제5권1호
• /
• pp.1-11
• /
• 2014

Lipids play important roles in biological systems; they store energy, play a structural role in the cell membrane, and are involved in cell growth, signal transduction, and apoptosis. Phospholipids (PLs) in particular have received attention in the medical and lipidomics research fields because of their involvement in human diseases such as diabetes, obesity, atherosclerosis, and many cancers associated with lipid metabolic disorders. Here I review experimental strategies for PL analysis based on nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (nLC-ESI-MSn). In particular, discussed are lipid extraction methods, nanoflow LC separation of PLs, effect of ionization modifiers on the ESI of PLs, influence of chain lengths and unsaturation degree of acyl chains of PLs on MS intensity, structural determination of the molecular structure of PLs and their oxidized products, and quantitative profiling of PLs from biological samples such as tissue, urine, and plasma in relation to cancer and coronary artery disease.

• BMB Reports
• /
• 제55권9호
• /
• pp.413-416
• /
• 2022

Ferroptosis is a type of programmed cell death distinct from apoptosis or necroptosis. Ferroptosis is well characterized by an iron-dependent accumulation of lipid peroxides and disruption of cellular membrane integrity. Many metabolic alterations can prevent or accelerate ferroptosis induction. Recent advances in analytical techniques of mass spectrometry have allowed high-throughput analysis of metabolites known to be critical for understanding ferroptosis regulatory metabolism. In this review, we introduce mass spectrometry-based analytical methods contributing to recent discovery of various metabolic pathways regulating ferroptosis, focusing on cysteine metabolism, antioxidant metabolism, and poly-unsaturated fatty acid metabolism.

• 한국영양학회:학술대회논문집
• /
• 한국영양학회 2003년도 추계학술대회 및 정기총회
• /
• pp.26-29
• /
• 2003

• 한국신재생에너지학회:학술대회논문집
• /
• 한국신재생에너지학회 2011년도 추계학술대회 초록집
• /
• pp.89.1-89.1
• /
• 2011

The influence of potential and humidity on the electrochemical carbon corrosion in high temperature polymer electrolyte membrane fuel cells(HT-PEMFCs) is investigated by measuring $CO_2$ emission at different potentials for 30 min using on-line mass spectrometry. These results are compared with low tempterature polymer electrolyte membrane fuel cells(LT-PEMFCs) operated at lower temperature and higher humidity condition. Although the HT-PEMFC is operated at non humidified condition, the emitted $CO_2$ in the condition of HT-PEMFC is more than LT-PEMFC at the same potential in carbon corrosion test. Thus, carbon corrosion shows a stronger positive correlation with the cell temperature. In addition, the presence of a little amount of water activate electrochemical carbon corrosion considerably in HT-PEMFC. With increased carbon corrosion, changes in fuel cell electrochemical characteristics become more noticeable and thereby indicate that such corrosion considerably affects fuel cell durability.

• Bulletin of the Korean Chemical Society
• /
• 제28권9호
• /
• pp.1499-1509
• /
• 2007

Proteomic analyses of synaptic vesicle fraction from rat brain have been performed for the better understanding of vesicle regulation and signal transmission. Two different approaches were applied to identify proteins in synaptic vesicle fraction. First, the isolated synaptic vesicle proteins were treated with trypsin, and the resulting peptides were analyzed using a high-pressure capillary reversed phase liquid chromatography/tandem mass spectrometry (cRPLC/MS/MS). Alternatively, proteins were separated by two-dimensional gel electrophoresis (2DE) and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF/MS). Total 18 and 52 proteins were identified from cRPLC/MS-MS and 2DE-MALDI-TOF-MS analysis, respectively. Among them only 2 proteins were identified by both methods. Of the proteins identified, 70% were soluble proteins and 30% were membrane proteins. They were categorized by their functions in vesicle trafficking and biogenesis, energy metabolism, signal transduction, transport and unknown functions. Among them, 27 proteins were not previously reported as synaptic proteins. The cellular functions of unknown proteins were estimated from the analysis of domain structure, expression profile and predicted interaction partners.

• 한국어병학회지
• /
• 제28권3호
• /
• pp.133-143
• /
• 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.