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Selection of substrate recognition sequence of protein kinase with ferric chelation affinity chromatography

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Science China(Life Sciences) 1997年第2期40卷 184-193页

作者：陈长征夏其昌李伯良王应睐 Shanghai Institute of Biochemistry Chinese Academy of Sciences Shanghai 200031 China

Protein kinase substrate phage (PKS phage) was constructed by fusing the substrate recognition consensus sequence of cAMP-dependent protein kinase (cAPK) with bacteriophage minor coat protein g3p and by dis-playing it on the surface of filamentous bacteriophage fd. Phosphorylation in vitro by cAPK showed a unique labelled band of approximately 60 ku, which was consistent with the molecular weight of the PKS-g3p fusion protein. Some weakly phosphorylated bands for both PKS phage and wild-type phage were also observed. Phage display random 15-mer peptide library phosphorylated by cAPK was selected with ferric (Fe3+ ) chelalion affinity resin. After 4 rounds of screening, phage clones were picked out to determine the displayed peptide sequences by DNA sequencing. The results showed that 5 of 14 sequenced phages displayed the cAPK recognition sequence motif (R)RXS/T. Their in vitro phosphorylation analyses revealed the specific labelled bands corresponding to the positive PKS phages with and without the typical (R)RXS/T sequence motif. It suggested that the new method of using ferric (Fe 3+ ) chelation affinity chromatography.to identify the substrate specificity of protein kinase from random peptide library was feasible.

关键词： protein kinase enzyme substrate phage display peptide library metal ion chelation affinity chromatography.

Multi-segment linear gradient optimization strategy based on resolution map in HPLC

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Science China Chemistry 2006年第4期49卷 315-325页

作者： Andreas Seidel-Morgenstern Max-Planck-Institut fr Dynamik Komplexer Technischer Systeme Magdeburg 39106 Germany

Based on the mechanism of chromatographic retention (the relationship between the retention of solute and the mobile phase conditions) and method of resolution map, several methods of optimizing multi-segment linear gradient elution conditions were proposed according to the different separation requirements of various samples. These methods were verified using literature data. Moreover, the advantages and disadvantages of these methods were compared. It was proved that the third method is a fast optimization method which is capable of separating all the components with relatively high resolution.

关键词： optimization, multi-segment, linear gradient, resolution map, liquid chromatography.

~1H-NMR studies on cis-trans isomers of spheroidenes

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Science China Chemistry 1996年第1期39卷 11-19页

作者：姜月顺小山泰 Department of Chemistry Jilin University Changchun 130023 China Faculty of Science Kwansei Gakuin University Nishinomiya 662 Japan

All-trans spheroidene was extracted from the cells of Rhodobacer sphaeroides 2.4.1 and a set of cis-trans isomers were isolated from an isomeric mixture obtained by iodine-sensitized photo-isomerization of the all-trans isomer by high-pressure liquid chromatography.(HPLC). On the basis of assignment of the 1H-NMR spectra (COSY, long-range COSY, rekyed COSY, ROESY and NOESY) of the all-trans isomer, the configurations of cis isomers were determined by the isomerization shifts of the olefinic 1H signals. The peaks of HPLC could be assigned as follows: A: 9,13’-cis, B1: 5,13-cis, B2: 13,9’-cis, C: 13-cis, E: 9-cis, F: 13’-cis, G: 5,9’-cis, H: 9’-cis, I: all-trans.

关键词： spheroidene cis-trans isomers 2D 1H-NMR high-pressure liquid chromatography.

Analysis of organic compounds in coal gasification wastewater

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Journal of Environmental Sciences 1992年第1期4卷 84-96页

作者： Wang Jusi and Zhao LihuiResearch Center for Eeo-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China

A procedure for analysis of organic pollutants in coal gasification wastewater was developed, including a series extraction steps at different pH, followed by LC separation or resin adsorption, then analyzed by GC or GC/MS. More than 200 organic pollutants in 22 categories were determined. CH2CL2 extraction at NaHCO3 presence was used to separate carboxylic acids with phenolic compounds in aqueous. Derivatization with acetic anhydride was used for analyses of mono-, di-, poly-hydroxyl phenolic compounds. 21 mono-hydroxyl phenols and 13 di-hydroxyl phenols were determined from the coai gasification wastewater samples. Derivatization with BF3-CH3OH was used for analysis of carboxylic acid. 17 mono-carboxyl, 4 di-carboxyl acids and 6 aromatic acids were determined from coal gasification wastewater samples.

关键词： wastewater analysis: coal gasification pollutants chromatography.

Nerve growth factor activity of several immunal related serine proteases

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Chinese Science Bulletin 1998年第22期43卷 1886-1891页

作者： Liying Guo Hong Zhu Yuancong Zhou Chinese Acad Sci Shanghai Inst Biochem Shanghai 200031 Peoples R China

Rabbit was immuned by the previously purified protein with high nerve growth factor (NGF) bioactivity (NGF_like protease) from \%Agkistrodon halys Pallas\% and the antisera were collected. The polyclonal antibodies were tentatively purified and then used as ligands of an affinity column. The \%***\% crude venom was fractionated by this affinity column and then by Mono Q on fast protein liquid chromatography.(FPLC). As a result, fraction Ⅱ and fraction Ⅲ were purified respectively, whose N_terminal amino acid sequences show high homology with the serine proteases in snake venoms, as well as the previous NGF_like protease. However, they possessed different levels of NGF bioactivity. The NGF activity of the previous NGF_like protease is equivalent to that of NGF, while the activity of fraction Ⅱ seems relatively low in contrast to fraction Ⅲ which had no NGF activity.

关键词： nerve growth factor neurotrophic factor serine protease affinity chromatography.

GC/ESR Studies on the Reactions of Fatty Alcohols With Activated Nitrogen in Microwave Discharge Processes

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Chinese Science Bulletin 1993年第17期38卷 1455-1459页

作者：陈德文王稳定陆道惠徐广智白守礼 Institute of Chemistry Academia Sinica Department of Applied Chemistry Beijing Institute of Chemical Engineering

Scientists have paid much attention to the plasma reaction of alcohol, especially of ethyl *** was known that N2 molecules in the microwave discharging processes can produce plasma and form several kinds of activated and excited species:Study on the reaction processes which are initiated by energy transfer from excited species to alcohol molecule is related not only to collision theory of reaction, but also to inter-molecular state-state reaction mechanism of excited species, therefore it is a very impor-

关键词： microwave discharge fatty alcohol ESR gas chromatography.

SYNTHESIS OF THE NOVEL CHIRAL POLYMERS WITH MULTIDENTATE LIGAND AND RESOLUTION OF DL-AMINO ACIDS

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Science China Chemistry 1989年第6期32卷 650-660页

作者：金仁华何炳林 Institute of Polymer Chemistry Nankai University Tianjin Department of Applied ChemistryBeijing Institute of Chemical Technology Institute of Polymer Chemistry Nankai University Tianjin

A new way for synthesizing styrenic type of chiral resins grafting to multidentate ligand has been found. The synthetic scheme is proved to be practical by means of elemental anaysis, IR spectra and metal adsorbility. The resolving power of the new chiral ligand resins coordinated with copper (Ⅱ) as stationary phases has been discussed based on HPLC, and some points of view about the compositions of mobil phase and mechanism of resolution are presented.

关键词： resolution ligand exchange enantioselectivity amino acids chromatography.

PREPARATION OF POLYVINYLAMINE LIGAND EXCHANGE RESIN GRAFTED WITH L-PROLIN

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Chinese Science Bulletin 1991年第11期36卷 903-906页

作者：袁直何炳林 Institute of Polymer Chemistry Nankai University Tianjin 300071 PRC

Chiral ligand polymers are the efficient stationary phases of liquid chromatography.for the resolution of DL-amino acids and are of great advantages for studying the mechanism of chiral recognition of the polymer. Up to now, the supporters used for resolution of DL-amino acids are based on polystyrene, polyacrylanide, polymethacrylate and polyvinylpyridine. We obtained a new chiral resin by grafting L-Pro on the polyvinylamine and then complexing it with Cu(Ⅱ) which could be used as a supporter of liquid chromatography.to resolve DL-Phe, His, Tryp. The factors affecting the functional reactions are also discussed.

关键词： chiral ligand exchange resin resolution liquid chromatography.

AFFINITY CHROMATOGRAPHIC PURIFICATION OF ACTIN FROM IMMATURE WHEAT ENDOSPERM AND PRELIMINARY STUDIES ON ITS CHARACTERISTICS

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Chinese Science Bulletin 1991年第3期36卷 235-238页

作者：金仕萍凌俊张伟成 Shanghai Institute of Plant Physiology Academia Sinica Shanghai 200032 PRC

In recent investigation, we have found that there is an active intercellular movement of protoplasm in the nuceUus and endosperm tissue of the developing wheat caryopses, which can be inversely arrested by cytochalasin B, suggesting that actin may be involved in this movement. So, the detection of the existence of actin in the tissues having

关键词： wheat endosperm actin affinity chromatography.