Accurate localization and relative quantification of arginine methylation using nanoflow liquid chromatography coupled to electron transfer dissociation and drbitrap mass spectrometry

Protein arginine (Arg) methylation serves an important functional role in eucaryotic cells, and typically occurs in domains consisting of multiple Arg in close proximity. Localization of methylarginine (MA) within Arg-rich domains poses a challenge for mass spectrometry (MS)-based methods; the peptides are highly charged under electrospray ionization (ESI), which limits the number of sequence-informative products produced by collision induced dissociation (CID), and loss of the labile methylation moieties during CID precludes effective fragmentation of the peptide backbone. Here the fragmentation behavior of Arg-rich peptides was investigated comprehensively using electron-transfer dissociation (ETD) and CID for both methylated and unmodified glycine-/Arg-rich peptides (GAR), derived from residues 679–695 of human nucleolin, which contains methylation motifs that are widely-represented in biological systems. ETD produced abundant information for sequencing and MA localization, whereas CID failed to provide credible identification for any available charge state (z=2–4). Nevertheless, CID produced characteristic neutral losses that can be employed to distinguish among different types of MA, as suggested by previous works and confirmed here with product ion scans of high accuracy/resolution by an LTQ/Orbitrap. To analyze MA-peptides in relatively complex mixtures, a method was developed that employs nano-LC coupled to alternating CID/ETD for peptide sequencing and MA localization/characterization, and an Orbitrap for accurate precursor measurement and relative quantification of MA-peptide stoichiometries. As proof of concept, GAR-peptides methylated in vitro by protein arginine N-methyltransferases PRMT1 and PRMT7 were analyzed. It was observed that PRMT1 generated a number of monomethylated (MMA) and asymmetric-dimethylated peptides, while PRMT7 produced predominantly MMA peptides and some symmetric-dimethylated peptides. This approach and the results may advance understanding of the actions of PRMTs and the functional significance of Arg methylation patterns.     

流动注射-分光光度法测定天然产物对酪氨酸酶活性的影响

建立了一种快速、灵敏且高精度的流动注射-分光光度法用于研究天然产物对酪氨酸酶活性的影响。此方法基于一定介质条件下L-3,4-二羟基苯丙氨酸(L-DOPA)在酪氨酸酶的作用下,发生氧化生成棕褐色多巴醌,其最大吸收峰位于475 nm处,而酪氨酸酶抑制剂能降低酶促反应速度,减少多巴醌形成的量,从而降低475 nm下的吸光度值,形成倒峰。在优化实验条件下,半抑制浓度的曲酸抑制酪氨酸酶活性测定相对标准偏差(n=10)为0.036%,与分光光度法和酶标仪微量法相比,精密度提高了10倍以上。咖啡酸对酪氨酸酶有较强的激活作用,对二酚酶相对激活率达到50%时的浓度(IC50)为0.170 mmol/L;薰衣草花水相提取物具有较强的抑制活性,其IC50值为0.96 mg/m L。与分光光度法及酶标仪微量法相比,流动注射-分光光度法精密度高,重复性好,适用于天然产物对酪氨酸酶活性影响的测定。     

果胶锆凝胶球对苯甲酸的吸附性能研究

用锆氧基离子与果胶反应制得果胶锆凝胶球,采用扫描电镜和红外光谱初步表征了凝胶球的结构,并测定了凝胶球的机械强度。研究了该凝胶球对苯甲酸的吸附性能。分别考察了果胶浓度、锆氧基离子浓度、吸附时间、p H值、温度及苯甲酸浓度对吸附性能的影响。结果表明,在298K下,果胶锆凝胶球对苯甲酸的吸附在4.5h左右达到平衡,当果胶的质量分数为3.0%,锆氧基离子质量分数为1.0%,苯甲酸初始浓度为500mg/L,吸附量可达73.89mg/g。所研究的吸附体系既适用于Freundlich方程,又适用于Langmuir方程;吸附过程为自发的放热、熵减过程,降低温度对吸附更有利。     

Synthesis and Ionic Conductivity of Network Polymer Electrolytes with Internal Plasticizers

Network polymer electrolytes with free oligo(oxyethylene) chains as internal plasticizers were prepared by cross-linking poly(ethylene glycol) acrylates. The effects of salt concentration and properties of internal plasticizers on ionic conductivity were studied.     

A Novel Preprocessing Algorithm for Three-Way HPLC Data

Orthogonal signal correction (OSC) was a data preprocessing algorithm. It ensured that the filtered information was irrelevant to concentration data while using it to filter the noise from the original data. This paper extended the OSC application range from two-way data to three-way data. Two drug data sets, Enoxacin, Norfloxacin, Ciprofloxacin and Betamethasone, cortisone acetate, prednisone acetate, showed that the application of the OSC algorithm to three-way HPLC data was feasible and needed further research.     

Selective oxidation of octadecan-1-ol to octadecanoic acid over Co3O4/SiO2 catalysts

Factors (reaction temperature, reaction time, flow rate of oxygen, amount of catalyst, etc.) influencing the catalytic properties of Co₃O₄/SiO₂catalyst in the oxidation octadecan-1-ol to octadecanoic acid were investigated. The catalysts were characterized by means of XRD, FT-IR and N₂-adsorption. The experimental results indicate that under the optimal condition the selectivity to octadecanoic acid reached 97.5 % over 5 % Co₃O₄/SiO₂ catalyst.This revised version was published online in December 2005 with corrections to the Cover Date.     

The effect of micro-morphology of Bi2O3 on catalytic properties of Co/Bi catalyst for wet air oxidation of acetic acid

The novel phase transfer catalysts S-8 [4-(dimethyloctylammonium) propansultan] and DB-X [1,4-bis(triethylmethylammonium)benzene dibromide] were synthesized and employed for high conversion synthesis of dichlorocyclopropane from various olefins.This revised version was published online in December 2005 with corrections to the Cover Date.     

Chiral separation and determination of ofloxacin enantiomers by ionic liquid-assisted ligand-exchange chromatography

A simple and accurate method for the separation and determination of ofloxacin enantiomers was developed by ionic liquid-assisted ligand-exchange high performance liquid chromatography. Both achiral and chiral ionic liquids were tested for their efficiency of ofloxacin enantiomeric separation. The effects of ligands, concentration of metal ion, organic modifier, pH of the mobile phase, and temperature were also investigated and evaluated. Optimal conditions were obtained on a conventional C(18) column, where the mobile phase consisted of methanol/water (20 : 80, v/v) (containing 4.0 mmol L(-1) amino acid ionic liquid and 3.0 mmol L(-1) copper sulfate) at a flow rate of 0.5 mL min(-1). Under this condition, the ofloxacin enantiomers could be baseline separated within 14 minutes; the proposed method was used to analyze different commercial ofloxacin medicines.     

Determination of the sodium 2-mercaptoethanesulfonate based on surface-enhanced Raman scattering

Based on the surface-enhanced Raman scattering (SERS) sodium 2-mercaptoethanesulfonate (mesna) was determined using unmodified gold colloid as the probe. The Raman scattering intensity was obviously enhanced in the presence of sodium chloride. The influence of experimental parameters, such as incubation time, sodium chloride concentration and pH value on SERS performance was examined. Under the optimum conditions, the SERS intensity is proportional to the concentration of mesna in the range of 9.0×10(-8) to 9.0×10(-7) mol/L and detection limit (S/N=3) is 1.16×10(-8) mol/L. The corresponding correlation coefficient of the linear equation is 0.996, which indicates that there is a good linear relationship between SERS intensity and mesna concentration. The experimental results indicate that the proposed method is a viable method for determination of mesna. The real samples were analyzed and the results obtained were satisfactory.     

金属酞菁化合物在TiO2表面的分散研究

Dispersion of copper(Ⅱ) phthalocyanine (CuPc), copper(Ⅱ) phthalocyaninesulfonate (CuPcS) and cobalt(Ⅱ)phthalocyaninetetrasulfonate (CoPcTS) on the surface of titanium dioxide was investigated by XRD, XPS, FT-IR and UV-Vis techniques. Results show that interaction between CuPc and TiO2 was very weak and CuPc was difficult to disperse on the surface of the support. While partly sulfurized CuPcS could be dispersed on the surface of support through sulfo-groups and its dispersion capacity was determined to be 0.085 g CuPcS/g TiO2. Completely sulfurlzed CoPcTS could also be dispersed on the surface of TiO2 as a monolayer and its dispersion capacity was 0.12 g CoPcTS/g TiO2. Interactions of the sulfo-groups as well as the electrons of CoPcTS with the surface of TiO2 could be evidenced by FT-IR characterization. Therefore, it was suggested that CoPcTS molecules be adsorbed on the surface of TiO2 in a flat-lying mode while CuPcS in a slanting one. UV-Vis spectra show that the dispersed CuPcS and CoPcTS molecules exist in both forms of monomers and dimers.