• Title/Summary/Keyword: fragment ions

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Altered APP Carboxyl-Terminal Processing Under Ferrous Iron Treatment in PC12 Cells

  • Kim, Chi Hyun;Yoo, Yeong-Min
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.3
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    • pp.189-195
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    • 2013
  • Amyloid-${\beta}$ peptide ($A{\beta}$), generated by proteolytic cleavage of the amyloid precursor protein (APP), plays a pivotal role in the pathogenesis of Alzheimer's disease (AD). The key step in the generation of $A{\beta}$ is cleavage of APP by beta-site APP-cleaving enzyme 1 (BACE1). Levels of BACE1 are increased in vulnerable regions of the AD brain, but the underlying mechanism is unknown. In the present study, we reported the effects of ferrous ions at subtoxic concentrations on the mRNA levels of BACE1 and a-disintegrin-and-metalloproteinase 10 (ADAM10) in PC12 cells and the cell responses to ferrous ions. The cell survival in PC12 cells significantly decreased with 0 to 0.3 mM $FeCl_2$, with 0.6 mM $FeCl_2$ treatment resulting in significant reductions by about 75%. 4,6-diamidino-2-phenylindole (DAPI) staining showed that the nuclei appeared fragmented in 0.2 and 0.3 mM $FeCl_2$. APP-${\alpha}$-carboxyl terminal fragment (APP-${\alpha}$-CTF) associations with ADAM10 and APP-${\beta}$-CTF with BACE1 were increased. Levels of ADAM10 and BACE1 mRNA increased in response to the concentrations of 0.25 mM, respectively. In addition, p-ERK and p-Bad (S112, S155) expressions were increased, suggesting that APP-CTF formation is related to ADAM10/ BACE1 expression. Levels of Bcl-2 protein were increased, but significant changes were not observed in the expression of Bax. These data suggest that ion-induced enhanced expression of AMDA10/BACE1 could be one of the causes for APP-${\alpha}/{\beta}$-CTF activation.

Confirmation of carbamate pesticides by liquid chromatography coupled with electrospray ionization mass spectrometry (LC/MS를 이용한 카바메이트계 잔류농약 확인방법)

  • 박준조;금모래;고광춘;박병옥;이성식
    • Korean Journal of Veterinary Service
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    • v.25 no.1
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    • pp.23-29
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    • 2002
  • A method was developed to separate, detect and qualify aldicarb, bendiocarb, carbaryl, carbofuran, ethiofencarb, methomyl, methiocarb, propoxur in meats and fruits. Experimental beef and fork samples were fortified with 0.05mg/kg of carbamate pesticides for analysis. Carbamate-detected pear by HPLC fluorescence detector(HPLC/FLS) are extracted with acetonitril and refined by solid phase extraction(SPE) filled with aminopropyl-bonded silca, In the following step, the injected materials into LC/MS are analyzed to result in the fact that bendiocarb, carbaryl, carbofuran, ethiofencarb, methomyl, methiocarb, propoxur presents several sorts of fraction ions following with; [M+H]$^{+}$, [M+Na]$^{+}$,[M-CONH$CH_3$]$^{+}$, [M-OCONH$CH_3$]$^{+}$. In addition, ethiofencarb presents [M-SCH$_2$$CH_3$]$^{+}$ ion distinctive and aldicarb presents [M+Na]$^{+}$ and [M-OCONH$CH_3$]$^{+}$ ion which is the most decisive fraction ion for pesticides such as bendiocarb, carbaryl, carbofuran, ethiofencarb, methiocarb, methomyl, propoxur excluding [M+H]$^{+}$ ion. However, [M-OCONH$CH_3$]$^{+}$ and [M-OCONH$CH_3$]$^{+}$ fraction ion charactering carbamate pesticides are detected most efficiently with fragment voltage 50ev. As a result, for rluantitative analysis, [M+Na]$^{+}$ ion is the most decisive ion for detection of aldicarb and [M+H]$^{+}$ ion is the most decisive fraction ion for Pesticides such as bendiocarb, carbaryl, carbofuran, ethiofencarb, methiocarb, methomyl, propoxur. Carbaryl-detected pear by HPLC/FLS are analyzed by L/MS and the result shows that [M+H]$^{+}$ and [M-CONH$CH_3$]$^{+}$ ions charactering carbaryl are detected.ering carbaryl are detected.

Analytical Methods of Levoglucosan, a Tracer for Cellulose in Biomass Burning, by Four Different Techniques

  • Bae, Min-Suk;Lee, Ji-Yi;Kim, Yong-Pyo;Oak, Min-Ho;Shin, Ju-Seon;Lee, Kwang-Yul;Lee, Hyun-Hee;Lee, Sun-Young;Kim, Young-Joon
    • Asian Journal of Atmospheric Environment
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    • v.6 no.1
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    • pp.53-66
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    • 2012
  • A comparison of analytical approaches for Levoglucosan ($C_6H_{10}O_5$, commonly formed from the pyrolysis of carbohydrates such as cellulose) and used for a molecular marker in biomass burning is made between the four different analytical systems. 1) Spectrothermography technique as the evaluation of thermograms of carbon using Elemental Carbon & Organic Carbon Analyzer, 2) mass spectrometry technique using Gas Chromatography/mass spectrometer (GC/MS), 3) Aerosol Mass Spectrometer (AMS) for the identification of the particle size distribution and chemical composition, and 4) two dimensional Gas Chromatography with Time of Flight mass spectrometry (GC${\times}$GC-TOFMS) for defining the signature of Levoglucosan in terms of chemical analytical process. First, a Spectrothermography, which is defined as the graphical representation of the carbon, can be measured as a function of temperature during the thermal separation process and spectrothermographic analysis. GC/MS can detect mass fragment ions of Levoglucosan characterized by its base peak at m/z 60, 73 in mass fragment-grams by methylation and m/z 217, 204 by trimethylsilylderivatives (TMS-derivatives). AMS can be used to analyze the base peak at m/z 60.021, 73.029 in mass fragment-grams with a multiple-peak Gaussian curve fit algorithm. In the analysis of TMS derivatives by GC${\times}$GC-TOFMS, it can detect m/z 73 as the base ion for the identification of Levoglucosan. It can also observe m/z 217 and 204 with existence of m/z 333. Although the ratios of m/z 217 and m/z 204 to the base ion (m/z 73) in the mass spectrum of GC${\times}$GC-TOFMS lower than those of GC/MS, Levoglucosan can be separated and characterized from D (-) +Ribose in the mixture of sugar compounds. At last, the environmental significance of Levoglucosan will be discussed with respect to the health effect to offer important opportunities for clinical and potential epidemiological research for reducing incidence of cardiovascular and respiratory diseases.

Structural Analysis of the Cu-binding Site in the [Cu·dCMP·dCMP-H]1- Complex

  • Jung, Sang-Mi;Kim, Ho-Tae
    • Mass Spectrometry Letters
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    • v.4 no.4
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    • pp.67-70
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    • 2013
  • The Cu-binding site in the $[Cu{\cdot}dCMP{\cdot}dCMP-H]^{1-}$ complex was investigated. The tandem mass (MS/MS) spectra of the [$[Cu{\cdot}dCMP{\cdot}dCMP-H]^{1-}$ parent ion showed $[dCMP{\cdot}Cu{\cdot}H_2PO_4+CONH]^{1-}$ fragment ions. Therefore, we propose that the Cu cation is simultaneously coordinated to the phosphate site and cytosine moiety in the stable geometry of the $[Cu{\cdot}dCMP{\cdot}dCMP-H]^{1-}$ complex. Three geometries for the complex were considered in an attempt to optimize the structure of the $[Cu{\cdot}dCMP{\cdot}dCMP-H]^{1-}$ complex. The ab initio calculations were performed at the $B3LYP/6-311G^{**}$ level.

Gamakamide C and D as Two New Analogues of Bitter-Tasting Cyclic Peptide with Hydantoin Structure from Oyster Crassostrea gigas

  • Jang, Jun Ho;Park, Taesung;Lee, Jong Soo
    • Fisheries and Aquatic Sciences
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    • v.18 no.2
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    • pp.131-135
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    • 2015
  • Two new bitter-tasting cyclic peptides comprising six amino acids, namely gamakamide C and D, were isolated from cultured oysters Crassostrea gigas. Dimethylaminoazobenzene sulfonyl-amino acid analysis detected Val and Leu in gamakamide C and Ile and Leu in gamakamide D. The molecular formula of gamakamide C was determined as $C_{43}H_{60}N_{7}O_8S$ by high-resolution fast atom bombardment mass spectroscopy (HR FAB-MS) ($[M+H]^+m/z822.4200{\Delta}-2.4mmu$), and that of gamakamide D was determined as $C_{43}H_{62}N_7O_8S$ by HR FAB-MS ($[M+H]^+m/z836.4379{\Delta}-2.0mmu$). Comparison of amino acid analyses and fragment ions by MS/MS among gamakamide C, D, and E (known), the structures of gamakamide C and D were confirmed $as-{\small{L}}-Val-{\small{L}}-Met(SO)-{\small{L}}-NMe-Phe-{\small{L}}-Leu-{\small{D}}-Lys-{\small{L}}-Phe-$ and $-{\small{L}}-Ile-{\small{L}}-Met(SO)-{\small{L}}-NMe-Phe-{\small{L}}-Leu-{\small{D}}-Lys-{\small{L}}-Phe-$, respectively.

Characterization of Low Molecular Weight Polyphenols from Pine (Pinus radiata) Bark

  • Mun, Sung-Phil;Ku, Chang-Sub
    • Food Science and Biotechnology
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    • v.15 no.3
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    • pp.424-430
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    • 2006
  • Low molecular weight polyphenols were isolated from hot water extracts of radiata pine (Pinus radiata) bark using a Sephadex LH-20 column and characterized by $^1H$ and $^{13}C$ NMR, UV, FT-IR, and GC-MS analyses. Major compounds isolated and identified were protocatechuic acid, trans-taxifolin, and quercetin. Trans-taxifolin, an important intermediate in biosynthetic route of proanthocyanidin (PA), was isolated in large quantities and indicates that PA is a major component of radiata pine bark. Small amounts of polyphenols were identified by GC-MS analysis. The presence of p-hydroxybenzoic acid, vanillic acid, protocatechuic acid, cis- and trans-feruic acid, p-coumaric acid, trans-caffeic acid, (-)-epicatechin, (+)-catechin, trans- and cis-taxifolin, (+)-gallocatechin, and quercetin was confirmed by comparison of mass fragmentation patterns and retention times (RT) with authentic samples. In addition, the presence of astringenin, astringenin glycoside, trans- and cis-leucodelphinidin was strongly assumed from characteristic mass fragment ions due to their conjugated structure and retro Diels-Alder reaction, and also from biosynthetic route of PA. GC-MS analysis allowed us to detect small amounts of phenolic acids and flavonoids and eventually discriminate trans- and cis-configuration in the identified polyphenols.

EF-hands in CBP7 are Important in the Process of Development

  • Dahyeon Kim;Taeck Joong Jeon;Byeonggyu Park;Dong Yeop SHIN
    • Journal of Integrative Natural Science
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    • v.17 no.1
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    • pp.31-41
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    • 2024
  • Calcium ions play an important role in development and intracellular signaling. Dictyostelium discoideum has 14 genes encoding calcium -binding proteins (CBPs), but the function of most CBPs during development has not yet been studied. In this study, we investigated the specific functions of CBP7, one of 14 CBPs, in development using RNA interference cell lines of CBP7, cell lines overexpressing CBP7, cell lines with point mutations in the EF-hand domain, and cell lines expressing fragment proteins. was intended to reveal. CBP7 consists of 169 amino acids and contains 4EF-hand domains. The CBP7-overexpressing cells showed complete loss of developmental process. These cells remained in the single-cell growth stage under development -inducing conditions, while wild-type cells formed aggregations within 6-8h of development and eventually formed fruiting bodies. The experiments using point-mutated CBP7 protein showed that all EF-hand domains of CBP7 were important for CBP7 to function during developmental process. These results suggest that CBP7 plays an important role in developmental processes across all EF-hand domains.

Synthesis and Crystal Structure of a New Quaternary Chalcoantimonide: KLa2Sb3S9 and KSm2Sb3Se8

  • Kim, Sung-Jin;Park, Sun-Ju;Yim, Sun-Ah
    • Bulletin of the Korean Chemical Society
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    • v.25 no.4
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    • pp.485-490
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    • 2004
  • Silver-needle shaped crystals of $KLa_2Sb_3S_9$ from $K_2S_x$ flux and $KSm_2Sb_3Se_8$ from NaCl/KCl flux reactions were obtained and their crystal structures were determined by the single crystal X-ray diffraction method. $KLa_2Sb_3S_9$ crystallizes in the orthorhombic noncentrosymmetric space group $P2_12_12_1$ (No.19) with a unit cell of a = 4.220(3) ${\AA}$, b = 24.145(2) ${\AA}$, c = 14.757(5) ${\AA}$ and Z = 4. $KSm_2Sb_3Se_8$ crystallizes in the orthorhombic space group Pnma (No.62) with a unit cell of a = 16.719(3) ${\AA}$, b = 4.1236(8) ${\AA}$, c = 22.151(4) ${\AA}$ and Z = 4. Both structures have three-dimensional tunnel frameworks filled with $K^+$ ions. $KSm_2Sb_3Se_8$ is an ordered version of $ALn_{1{\pm}X}B_i{4{\pm}X}S_8$, and it is made up of NaCl-type and $Gd_2S_3$-type fragments. $KLa_2Sb_3S_9$ also contains building fragments similar to those of $KSm_2Sb_3Se_8$, however, there are chalcogen-chalcogen bonds in the $Gd_2S_3$-type fragment. The formula of $KLa_2Sb_3S_9$ can be described as $(K^+ )(La^{3+})_2(Sb^{3+})^3(S^{2-})_7(S_2^{2-})$.

Study on analytical method of residual benzimidazole anthelmintics in meat by LC/MS (LC/MS를 이용한 식육중 잔류 벤지미다졸계 구충제 분석법 연구)

  • Choi Eun-Young;Seo Heyng-Seok;Baek Kui-Jeong;Hur Boo-Hong;Seo Lee-Won;Joung Dong-Suk
    • Korean Journal of Veterinary Service
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    • v.28 no.1
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    • pp.81-89
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    • 2005
  • Recently, mass spectrometry coupled with liquid chromatography (LC/MS) has been a preferred technique for determination of organic compounds in complex matrixes. LC/MS provides a high degree sensitivity and specificity of the compounds of interest. The purpose of this study was to confirm analytical method of residual 6 benzimidazoles (thiabendazole, oxfendazole, mebendazole, albendazole, flubendazole and fenbendazole) in meat by LC/MS. Benzimidazoles were analyzed by LC/MS on XTerra $C_{18}$ column with 0.01% trifluoroacetic acid-acetonitrile (TFA) in a gradient mode as mobile phase, and that were identified by electrospray ionization with selected ion recording mode at 150-350 amu mass range. Residual benzimidazoles were extracted from tissue with ethylacetate, and elute benzimidazoles with $50\%$ acetonitrile. In the LC/MS analysis of benzimidazoles, signal to noise ratio was showed relatively high in the positive mode and special ion in the quality analysis was determined via $[M+H]^+$ and Fragment ions. A spectrum of benzimidazoles was showed from all 6 benzimidazoles

Purification and Biochemical Characterization of a Novel Fibrinolytic Enzyme from Streptomyces sp. P3

  • Cheng, Guangyan;He, Liying;Sun, Zhibin;Cui, Zhongli;Du, Yingxiang;Kong, Yi
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1449-1459
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    • 2015
  • A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg2+ and Ca2+ and completely inhibited by Cu2+, but slightly enhanced by Fe2+. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.