• BMB Reports
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• v.47 no.10
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• pp.593-598
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• 2014

RNA polymerase II carboxyl-terminal domain (RNAPII CTD) phosphatases are responsible for the dephosphorylation of the C-terminal domain of the small subunit of RNAPII in eukaryotes. Recently, we demonstrated the identification of several interacting partners with human small CTD phosphatase1 (hSCP1) and the substrate specificity to delineate an appearance of the dephosphorylation catalyzed by SCP1. In this study, using the established cells for inducibly expressing hSCP1 proteins, we monitored the modification of ${\beta}$-O-linked N-acetylglucosamine (O-GlcNAc). O-GlcNAcylation is one of the most common post-translational modifications (PTMs). To gain insight into the PTM of hSCP1, we used the Western blot, immunoprecipitation, succinylayed wheat germ agglutinin-precipitation, liquid chromatography-mass spectrometry analyses, and site-directed mutagenesis and identified the $Ser^{41}$ residue of hSCP1 as the O-GlcNAc modification site. These results suggest that hSCP1 may be an O-GlcNAcylated protein in vivo, and its N-terminus may function a possible role in the PTM, providing a scaffold for binding the protein(s).

• The Korean Journal of Community Living Science
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• v.24 no.1
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• pp.5-12
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• 2013

Caffeine is an alkaloid of the methylxanthine family known as a central nervous system stimulant, temporarily warding off drowsiness and restoring alertness in humans. There is a recommended upper limits of caffeine for health because a high dose can cause negative effects. Chlorogenic acid is a natural polyphenol compound known to have an antioxidant activity. In this study, the contents of caffeine and chlorogenic acid in coffee beans from different origins(Costa Rica, Indonesia, Vietnam) were determined by using liquid chromatography-tandem mass spectrometry(LC-MS/MS). The experiment offers more selectivity and sensitivity for those compounds compared with conventional methods such as UV/VIS spectrophotometry. The average concentrations of caffeine and chlorogenic acid in coffee beans origined in Costa Rica were 15.05 mg/g and 5.33 mg/g respectively. In the case of coffee beans origined in Indonesia, the average concentrations were 13.10 mg/g for caffeine and 3.75 mg/g for chlorogenic acid. Vietnamese coffee showed that the average concentrations were 17.79 mg/g for caffeine and 1.12 mg/g for chlorogenic acid. This study can contribute to a better understanding of the contents of caffeine and chlorogenic acid in various coffee beans in order to evaluate dietary intake.

• Korean Journal of Pharmacognosy
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• v.45 no.2
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• pp.141-146
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• 2014

In this study, we investigated on antioxidative activities and tyrosinase inhibitory activities of methanol extract and its fractions from roots of Arctium lappa. The total phenolic compound and flavonoid content of the ethylacetate fraction was found to be 818.29 mg/g and 360.59 mg/g as the highest content. In the measurement of DPPH radical scavenging ability and tyrosinase inhibitory activity, the ethylacetate fraction was higher than the other fractions and the extract. In addition, comparative analysis of phenolic compounds by liquid chromatography-mass spectrometry (MS)/MS system under the multiple-reaction monitoring (MRM) with negative-ion electrospray ionization mode. The main phenolic compounds in the extract and fractions of roots from Arctium lappa were cynarin and chlorogenic acid. The main phenolic compound of the ethylacetate fraction was cynarin. n-Butanol fraction had a significantly higher chlorogenic acid content than other samples. In conclusion, DPPH radical scavenging ability and tyrosinase inhibitory activity of the cynarin-riched ethylacetate fraction showed the highest activity.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.13
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• pp.5417-5421
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• 2014

Objectives: The identification of cutaneous T cell lymphoma (CTCL) biomarkers may serve as a predictor of disease progression and treatment response. The aim of this study was to map potential biomarkers in CTCL plasma. Design and Methods: Plasma metabolic perturbations between CTCL cases and healthy individuals were investigated using metabolomics and ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Results: Principal component analysis (PCA) of the spectra showed clear metabolic changes between the two groups. Thirty six potential biomarkers associated with CTCL were found. Conclusions: Based on PCA, several biomarkers were determined and further identified by LC/MS/MS analysis. All of these could be potential early markers of CTCL. In addition, we established that heparin as a nticoagulant has better pre-treatment results than EDTA with the UHPLC-QTOF/MS appraoch.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.2
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• pp.129-134
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• 2014

Two neuropeptides were purified from the acidified tube feet extract of the starfish Asterias amurensis by C18 reversed phase and size-exclusion high-performance liquid chromatography (HPLC). The tube feet extract and the purified peptides (AST-I and AST-II) showed potent contractile activity on dorsal retractor muscle (DRM) of the starfish Asterina pectinifera and intestine (smooth muscle) of the panther puffer Takifugu pardalis. Treatment of the purified peptides with dithiothreitol (DTT) for 60 min at $37^{\circ}C$ significantly altered their retention times, suggesting that these compounds contained disulfide bonds. The molecular weights of AST-I and AST-II were determined to be 2064.2 Da and 6137.2 Da, respectively, by MALDI-TOF mass spectrometry.

• Clinical and Experimental Pediatrics
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• v.59 no.sup1
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• pp.41-44
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• 2016

We report here a case of maternal 3-methylcrotonyl-coenzyme A carboxylase (3-MCC) deficiency in a Korean woman. Her 2 infants had elevated 3-hydroxyisovalerylcarnitine (C5-OH) on a neonatal screening test by liquid chromatography-tandem mass spectrometry (LC-MS/MS), but normal results were found on urine organic acid analysis. The patient was subjected to serial testing and we confirmed a maternal 3-MCC deficiency by blood spot and breast milk spot test by LC-MS/MS, serum amino acid analysis, urine organic acid and molecular genetic analysis that found c.838G>T (p.Asp280Tyr) homozygous mutation within exon 9 of the MCCB gene. Especially, we confirmed marked higher levels of C5-OH on breast milk spot by LC-MS/MS, in the case of maternal 3-MCC deficiency vs. controls.

• Fisheries and Aquatic Sciences
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• v.18 no.2
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• pp.165-171
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• 2015

Abalone containing phlorotannins is produced by feeding the phlorotannin-rich brown seaweed Eisenia bicyclis after 4 days of starvation. Reverse-phase high-performance liquid chromatography (RP-HPLC) was used to isolate and quantify phlorotannins, which were identified by mass spectrometry and [$^1H$]-nuclear magnetic resonance to be the P1 compound, 7-phloroeckol, and eckol. When E. bicyclis was used as feed, P1 compound accumulated to an average of 1.60 mg/g dry weight of abalone muscle tissue after 18 d, 7-phloroeckolol to 0.21 mg/g after 16 d, and eckol to 0.22 mg/g after 12 d. Saccharina japonica was used as a control feed, and the abalone showed little or no accumulation of phlorotannins in muscle tissue. Feed consumption and growth rate were very similar when either E. bicyclis or S. japonica was fed for 20 d. Half-maximal reductions in the levels of P1 compound, 7-phloroeckol, and eckol accumulation were attained in 1.5, 1.9, and 3.4 days, respectively, after the feed was switched from E. bicyclis to S. japonica. Value-added abalone containing bioactive phlorotannins can be produced by simply changing the feed to the phlorotannin-rich E. bicyclis 18 d prior to harvesting.

• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2127-2137
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• 2016

A mangrove microbial community was analyzed at the gene and protein levels using metagenomic and proteomic methods with the green macroalgae Enteromorpha prolifera as the substrate. Total DNA was sequenced on the Illumina HiSeq 2000 PE-100 platform. Two-dimensional gel electrophoresis in combination with liquid chromatography tandem mass spectrometry was used for proteomic analysis. The metagenomic data revealed that the orders Pseudomonadales, Rhizobiales, and Sphingomonadales were the most prevalent in the mangrove microbial community. By monitoring changes at the functional level, proteomic analyses detected ATP synthase and transporter proteins, which were expressed mainly by members of the phyla Proteobacteria and Bacteroidetes. Members of the phylum Proteobacteria expressed a high number of sugar transporters and demonstrated specialized and efficient digestion of various glycans. A few glycoside hydrolases were detected in members of the phylum Firmicutes, which appeared to be the main cellulose-degrading bacteria. This is the first report of multiple "omics" analysis of E. prolifera degradation. These results support the fact that key enzymes of glycoside hydrolase family were expressed in large quantities, indicating the high metabolic activity of the community.

• Korean Journal of Pharmacognosy
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• v.49 no.1
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• pp.70-75
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• 2018

Acer tegmentosum Maxim (ATM) has been used to treat hepatic disorders in traditional oriental medicine. However, there is little information about phytochemical constituents for quality control of ATM. In this study, we developed and established a simultaneous analytical method of the 10 marker compounds (three coumarins, 3 flavonoids, 1 lignan, 3 phenolics) in ATM using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS). Chromatographic separation of ten target analytes was achieved with a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$), using a mobile phase of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. Identifications and quantitation of all analytes were performed using a Q-Exactive UPLC-MS/MS system. Correlation coefficients of the calibration curve for all analytes were ${\geq}0.9986$. The values of limits of detection and quantification of all analytes were 0.5-10.0 and 5.0-50.0 ng/mL, respectively. The established UPLC-MS/MS method successfully identified all target analytes in ATM, and the phytochemicals were 0.01-67.98 mg/g in its lyophilized water extract.

• Korean Journal of Veterinary Research
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• v.50 no.1
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• pp.11-17
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• 2010

Linear and non-linear regressions were used to derive the calibration function for the measurement of roxithromycin plasma concentration. Their results were compared with weighted least squares regression by usual weight factors. In this paper the performance of a non-linear calibration equation with the capacity to account empirically for the curvature, y = ax$^{b}$ + c (b $\neq$ 1) is compared with the commonly used linear equation, y = ax + b, as well as the quadratic equation, y = ax$^{2}$+ bx + c. In the calibration curve (range of 0.01 to 10 ${\mu}g/mL$) of roxithromycin, both heteroscedasticity and nonlinearity were present therefore linear least squares regression methods could result in large errors in the determination of roxithromycin concentration. By the non-linear and weighted least squares regression, the accuracy of the analytical method was improved at the lower end of the calibration curve. This study suggests that the non-linear calibration equation should be considered when a curve is required to be fitted to low dose calibration data which exhibit slight curvature.