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Display of aggregation-prone ligand binding domain of human PPAR gamma on surface of bacteriophage lambda

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Acta Pharmacologica Sinica 2006年第1期27卷 91-99页

作者： Bo KONG~(2,4) Wei-jun MA~(2,3,5)2 institute of Health sciences.shanghai institutes for Biological sciences,chinese academy of sciences and medical School of shanghai Jiao Tong University.shanghai 200025.china 3 Shaughai Research Center of Biotechnology,shanghai institutes for Biological sciences,Chiese academy og sciences.shanghai 200233.china 4 Graduate School of the chinese academy of sciences instituteofHealthsciences ShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesandMedicalSchoolofShanghaiJiaoTongUniversityShanghai200025China

Aim:To display the aggregation-prone ligand binding domain (LBD) of the hu-man peroxisome proliferator-activated receptor gamma (PPARγ) on the surface ofbacteriophages to establish an easy screening assay for the identification of PPARγ***:Plasmids were constructed for the expression of the PPARγLBD as a fusion to the N-terminus of the g3p protein of filamentous phage or theC-terminus of the capsid protein D (pD) of phage *** fusion proteinswere expressed in E coli and solubility characteristics were *** against the LBD as well as the pD protein were prepared for Westernblot analysis and phage capture ***:The pD-LBD fusion protein waspartially soluble,whereas the LBD-g3p fusion protein was detected only in theinsoluble *** pD-LBD fusion protein was efficiently incorporated inphage ***,the LBD was shown to be displayed on the surfaceof bacteriophage *** average,the pD-LBD fusion protein accounted for28% of the total pD protein in the lambda head ***:The hydro-phobic PPARγ LBD was expressed as a soluble form of fusion protein in E coli anddisplayed on the surface of bacteriophage lambda when it was fused to the lambdapD *** lambda pD fusion system could be used for improving the solu-bility of proteins that tend to form inclusion bodies when expressed in E *** phage particles displaying the LBD of PPARy may be of great value for theidentification of novel PPARγ ligands.

关键词： protein expression lambda phage surface display nuclear receptor peroxisome proliferator-activated receptors ligand binding domain

Design,synthesis,and anti-inflammatory evaluation of a series of novel amino acid-binding 1,5-diarylpyrazole derivatives

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Acta Pharmacologica Sinica 2005年第7期26卷 865-872页

作者： Ming-hui LI Lin-lin YIN Mao-jun CAI Wei-yu ZHANG Yue HUANG Xin WANG Xing-zu ZHU~1 Jing-kang SHEN~1 State Key Laboratory of Drug Research,shanghai institute of materia medica,shanghai institutes for Biological sciences,chinese academy of sciences,Graduate School of the chinese academy of sciences.shanghai 201203,china State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai 201203 China

Aim:To design and synthesize a series of novel amino acid-binding 1,5-diarylpyrazole derivatives,which are intended to act as prodrugs with better aque-ous solubility than celecoxib,and which will exert potent anti-inflammatory activi-ties after being converted to their parent compounds in ***:To intro-duce an amino acid,celecoxib analogs containing amino or methylamino groupwere synthesized first through multi-step chemical *** the synthesizedcompounds were screened in an intact cell-based assay in vitro and in carrag-eenan-induced mouse paw edema in *** active compounds were selectedfor further evaluation in a carrageenan-induced rat paw edema *** prelimi-nary pharmacokinetics experiments were conducted using high performance liq-uid chromatography/mass spectrometry(HPLC/MS).Results:Celecoxib,6 of the1,5-diarylpyrazole class of celecoxib analogs,and their amino acid derivatives(hydrochloride salts)were *** vitro screening,the hydrochloridesalts showed decreased inhibitory effects on cyclooxygenase(COX)-1 and COX-2 compared with their parent compounds,but some exhibited potent anti-inflam-matory activity in *** 4a was selected for further evaluation,and itsanti-inflammatory effect was equivalent to that of celecoxib after oral administra-tion in the carrageenan-induced rat paw edema *** three doses(25 mg/kg,50 mg/kg,and 100 mg/kg)the percentage inhibition on edema was 20.7%,52.6%,and 62.6%(for compound 4a)and 27.8%,38.4%,and 40.1%(for celecoxib),*** pharmacokinetic evaluations support the hypothesisthat compound 4a was actually converted to its parent compound,compound ***:The compound bound with amino acid acts like prodrug,which canexert anti-inflammatory effect similar to celecoxib after being converted to its par-ent *** finding will be of great benefit in carrying out structuralmodifications of prodrug-like selective COX-2 inhibitors.

关键词： nonsteroidal anti-inflammatory agents cyclo oxygenase inhibitors celecoxib pyrazole sulfonamide prodrugs

Kinetic Process and Conductivity of Ag^+ Ion Exchange for Polycrystalline Na-β"-Al_2O_3

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Journal of materials Science & Technology 1996年第4期12卷 315-317页

作者： Jianhua YANG(shanghai institute of Ceramics, chinese academy of Scieoces, shanghai 200050, china) shanghai institute of Ceramics Chinese Academy of Sciences Shanghai 200050 China

This paper reports the kinetic process of Ag ion exchange for the polycrystalline Na-β'-*** interdiffusion coefficients in the process of Ag+ and Na+ ion exchange have been calculated with an 'one dimensional double side diffusion model'. Microstructures of the samples were observed and analysed by XRD, EMPA, SEM. The results of the conductivity measurements for samples with Na+, Ag+ and Na+-Ag+ mobile ions are presented and explained

关键词： Kinetic Process and Conductivity of Ag Ion Exchange for Polycrystalline Na Al2O3

Expression of Monomeric Insulin Precursor in Pichia pastoris and Its Conversion into Monomeric B27 Lys Destripeptide Insulin by Tryptic Hydrolysis

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Acta Biochimica et Biophysica Sinica 2005年第4期37卷 234-240页

作者： Jin-Guo DING~(1,3) Jian FEI~2 Da-Fu CUI~1 You-Shang ZHANG~1 ~1institute of Biochemistry and Cell Biology,and ~2Model Organism Research Center,shanghai institutes for Biologicol sciences, chinese academy of sciences.shanghai 200031,china ~3Graduate School of the chinese academy of sciences.shanghai 200031,china institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China Graduate School of the chinese academy of sciences.shanghai 200031 China Model Organism Research Center Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Monomeric B27 Lys destripeptide insulin (B27 Lys DTrI) was designed and produced from itsprecursor expressed in Pichia pastoris through tryptic hydrolysis instead of the less efficient *** monomeric B27 Lys DTrI precursor (MIP) was purified from a cultured medium *** by a combination of hydrophobic,size-exclusion,and ion-exchange *** purifiedMIP was converted,by tryptic hydrolysis,to B27 Lys DTrI,which was then purified by ion-exchangechromatography to homogeneity as assessed by native gel electrophoresis,HPLC,amino acid composition,and electrospray mass-spectrometric analysis.B27 Lys DTrI exhibited superior monomeric properties insize-exclusion *** yield of MIP was 200 mg per liter of culture,and the overall yield ofpurified B27 Lys DTrI from the crude MIP was 70%.The in vivo biological activity of B27 Lys DTrI asdetermined by the mouse convulsion assay was 21 U/mg,identical to that obtained by semisynthesis.

关键词： monomeric B27 Lys destripeptide insulin tryptic hydrolysis Pichia pastoris

Positive Charges on Lysine Residues of the Extrinsic 18 kDa Protein Are Important to Its Electrostatic Interaction with Spinach Photosystem Ⅱ Membranes

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Acta Biochimica et Biophysica Sinica 2005年第11期37卷 737-742页

作者： Jin-Peng GAO Zhen-Hua YONG Feng ZHANG Kang-Cheng RUAN Chun-He XU Gen-Yun CHEN institute of Plant Physiology & Ecology.shanghai institutes for Biological sciences,chinese academy of sciences.shanghai 200032,china Laboratory of Proteomics,institute of Biochemistry and Cell Biology,shanghai institutes for Biological sciences,chinese academy of sciences.shanghai 200031,china College of sciences,Zhejiang University.Hangzhou 310029,china Graduate School of the chinese academy of sciences.shanghai 200032,china institute of Plant Physiology & Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China Graduate School of the chinese academy of sciencesshanghai 200032 China College of sciences Zhejiang UniversityHangzhou 310029 China Laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China

To determine the contribution of charged amino acids to binding with the photosystem Ⅱcomplex(PSⅡ),the amino or carboxyl groups of the extrinsic 18 kDa protein were modified with N-succinimidyl propionate(NSP)or glycine methyl ester(GME)in the presence of a water-soluble carbodiimide,*** on isoelectric point shift,4-10 and 10-14 amino groups were modified in the presence of2 and 4 mM NSP,***,3-4 carboxyl groups were modified by reaction with *** of negatively charged carboxyl groups with GME did not alter the binding activity of theextrinsic 18 kDa ***,the NSP-modified 18 kDa protein,in which the positively charged aminogroups had been modified to uncharged methyl esters,failed to bind with the PSⅡ membrane in the presenceof the extrinsic 23 kDa *** defect can not be attributed to structural or conformational alterationsimposed by chemical modification,as the fluorescence and circular dichroism spectra among native,GME-and NSP-modified extrinsic 18 kDa proteins were ***,we have concluded that the positive chargesof lysyl residues in the extrinsic 18 kDa protein are important for its interaction with PSⅡ membranes in thepresence of the extrinsic 23 kDa ***,it was found that the negative charges of carboxylgroups of this protein did not participate in binding with the extrinsic 23 kDa protein associated with PSⅡmembranes.

关键词： extrinsic 18 kDa protein circular dichroism electrostatic interaction chemical modification

Tetrahydroacridine inhibits voltage-dependent Na~+ current in guinea-pig ventricular myocytes

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Acta Pharmacologica Sinica 2004年第9期 36-42页

作者： Wei WANG, Yi-ping WANG, Guo-yuan HUState Key Laboratory of Drug Research, The First Department of Pharmacology, shanghai institute of materia medica, shanghai institutes for Biological sciences, chinese academy of sciences, shanghai 201203, china

AIM: To study the effects of tetrahydroacridine (tacrine) on voltage-gated Na+ channels in cardiac tissues. METHODS: Single ventricular myocytes were enzymatically dissociated from adult guinea-pig heart. Voltage-depe... 详细信息

关键词： Alzheimer disease cholinesterase inhibitors sodium channels tetrahydroacridine (tacrine) myocardium patch-clamp techniques

Possible Role of Histone H1 in the Regulation of Furin-dependent Proprotein Processing

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Acta Biochimica et Biophysica Sinica 2007年第3期39卷 173-180页

作者： Jinbo HAN~1 Jianxin GU~2 Chengwu CHI~(1,3*) 1 institute of Biochemistry and Cell Biology,chinese academy of sciences,Graduate School of the chinese academy of sciences.shanghai 200031,china 2 Gene Research Center,medical Center of Fudan University,shanghai 200032,china 3 institute of Protein Research,Tongji University,shanghai 200092,china institute of Biochemistry and Cell Biology Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai China.

Histone H1 and its C-terminal lysine rich fragments were recently found to be potent inhibitorsof furin,a mammalian proprotein ***,its role in the regulation of furin-dependent proproteinprocessing remains *** we report that histone H1 efficiently blocks furin-dependent pro-yonWillebrand factor（pro-vWF）processing in a dose-dependent *** and immunof-luorescence studies confirmed that histone H1 could interact with furin,and the interaction mainly took placeon the cell *** noted that histone H1 was released from cells undergoing necrosis and apoptosisinduced by *** findings suggested that histone H1 might be involved in extracellular and/or intracellu-lar furin regulation.

关键词： furin histone H1 pro-vWF proprotein processing

Axon guidance and neuronal migration research in china

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Science china(Life sciences) 2010年第3期53卷 304-314页

作者： YUAN XiaoBing institute of Neuroscience,shanghai institutes for Biological sciences,chinese academy of sciences.shanghai 200031,china institute of Neuroscience Shanghai Institutes for Biological Sciences Chinese Academy of Sciences

Proper migration of neuronal somas and axonal growth cones to designated locations in the developing brain is essential for the assembly of functional neuronal *** progress in research of axon guidance and neuronal migration has been made in the last twenty *** researchers began their exploration in this field ten years ago and have made significant contributions in clarifying the signal transduction of axon guidance and neuronal *** unique experimental approaches,including the migration assay of single isolated neurons in response to locally delivered guidance cues,have been developed by chinese neuroscientists to investigate the molecular machinery underlying these guidance events.

关键词： axon guidance neuronal migration development neuronal circuits

A New Flavan-3-ol Glucoside from Daphniphyllum oldhami

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chinese Chemical Letters 2003年第9期14卷 926-929页

作者： Zhi Yu SHAO, Da Yuan ZHU, Yue Wei GUOState Key Laboratory of Drug Research, institute of materia medica, shanghai institutes for Biological sciences, chinese academy of sciences, shanghai 200031 State Key Lab. of Drug Research Shanghai Institutes for Biol. Sci. Chinese Academy of Sciences Shanghai 200031 China

A new flavan-3-ol glucoside, (-)-afzelechin-7-O-β-D-glucopyranoside (1), has been isolated from the stem of Daphniphyllum oldhami. The structure of the new compound was elucidated on the basis of detailed spectroscop... 详细信息

关键词： Daphniphyllum oldhami flavan-3-ol (-)-afzelechin-7-O-β-D-glucopyranoside.